Sem10Biol Mitocondriopatias

SEMINARIO SEMANA N 10 BIOLOGIA CELULAR Y MOLECULAR FACULTAD DE MEDICINA HUMANA
ADN MITOCONDRIAL. APLICACIONES EN MEDICINA FORENSE. MITOCONDRIOPATAS

1) Mencione las diferencias estructurales y funcionales que existen entre la membrana mitocondrial interna y externa. 2) Por qu se dice que las mitocondrias son estructuras dinmicas? 3) Qu ocurre con los componentes mitocondriales despus de la fusin? 4) Qu enfermedades son producidas por mutaciones en genes esenciales para la dinmica mitocondrial? 5) ADN mitocondrial y ADN nuclear: semejanzas, diferencias y cmo se heredan. 6) Mitocondriopatas: Caractersticas comunes, clasificacin. Investigue las bases moleculares de dos enfermedades producidas por estos defectos. 7) Por qu es importante la matriz mitocondrial? 8) Si existe algn dao en las mitocondrias de una persona cul de los siguientes procesos se veran directamente afectados?, explique brevemente cada uno de los procesos afectados: a. Glucolisis. b. Ciclo de Krebs. c. Transporte de electrones. d. Fosforilacin oxidativa. e. Fermentacin. 9) Qu utilidad tiene el ADN mitocondrial en la medicina forense? Mencione algunas aplicaciones.
Lecturas: Detmer S and Chan D. Functions and dysfunctions of mitochondrial dynamics. Nature Reviews Mol Cell Bio 2007; 8: 870-879. Solano A, Playn A, Lpez-Prez M y Montoya M. Enfermedades genticas del ADN mitocondrial humano. Salud Pblica de Mxico 2001; 43:151-161.
REVIEWS
Functions and dysfunctions of mitochondrial dynamics

Scott A. Detmer and David C. Chan
Abstract | Recent findings have sparked renewed appreciation for the remarkably dynamic nature of mitochondria. These organelles constantly fuse and divide, and are actively transported to specific subcellular locations. These dynamic processes are essential for mammalian development, and defects lead to neurodegenerative disease. But what are the molecular mechanisms that control mitochondrial dynamics, and why are they important for mitochondrial function? We review these issues and explore how defects in mitochondrial dynamics might cause neuronal disease.
Cristae
Invaginationsofthe mitochondrialinner membrane.
Nebenkern structure
Acytosolicstructure,foundin someinsectspermatids,thatis formedbythefusionof mitochondria.
Division of Biology, California Institute of Technology, Pasadena, California 91125, USA. Correspondence to D.C.C. e-mail: dchan@caltech.edu doi:10.1038/nrm2275 Publishedonline 10October2007
In the 1950s, seminal electron microscopy studies led to the canonical view of mitochondria as bean-shaped organelles. These studies revealed the ultrastructural hallmarks of mitochondria, which include double lipid membranes and unusual inner membrane folds termed cristae . Recent studies have led to renewed appreciation for the fact that the mitochondrial structure is highly dynamic1,2. Mitochondria have drastically different morphologies depending on the cell type and, even in the same cell, mitochondria can take on a range of morphologies, from small spheres or short rods to long tubules. In fibroblasts, for example, mitochondria visualized with fluorescent proteins or specific dyes typically form tubules with diameters of ~0.5 mm, but their lengths can range from 110 mm or more. Even more remarkably, imaging studies in live cells indicate that mitochondria constantly move and undergo structural transitions. Mitochondrial tubules move with their long axes aligned along cytoskeletal tracks3. Individual mitochondria can encounter each other during these movements and undergo fusion, resulting in the merging of the double membranes and the mixing of both lipid membranes and intramitochondrial content (BOX1). Conversely, an individual mitochondrion can divide by fission to yield two or more shorter mitochondria. What are the molecular mechanisms that underlie these unusual behaviours, and do they have consequences for mitochondrial function and cell physiology? In this Review, we discuss the dynamic nature of mitochondria and summarize the mechanisms that drive mitochondrial fusion and fission. In addition, we discuss recent insights into how these processes affect the function of mitochondria. As well as controlling the
shape of mitochondria, fusion and fission are crucial for maintaining the functional properties of the mitochondrial population, including its respiratory capacity. Moreover, mitochondrial dynamics has key roles in mammalian development, several neurodegenerative diseases and apoptosis.
Mitochondria as dynamic organelles By several criteria, mitochondria are dynamic organelles. First, the shape and size of mitochondria are highly variable and are controlled by fusion and fission. Second, mitochondria are actively transported in cells and they can have defined subcellular distributions. Finally, the internal structure of mitochondria can change in response to their physiological state.
Dynamic shape. The length, shape, size and number of mitochondria are controlled by fusion and fission (FIG.1a). At steady state, the frequencies of fusion and fission events are balanced 4 to maintain the overall morphology of the mitochondrial population. When this balance is experimentally perturbed, dramatic transitions in mitochondrial shape can occur. Genetic studies in yeast and mammals indicate that cells with a high fusion-to-fission ratio have few mitochondria, and that these mitochondria are long and highly interconnected58 (FIG.2). Conversely, cells with a low fusionto-fission ratio have numerous mitochondria that are small spheres or short rods these are often referred to as fragmented mitochondria. In vivo, such changes in dynamics control mitochondrial morphology during development. For example, during Drosophila melanogaster spermatogenesis, many mitochondria synchronously fuse to form the Nebenkernstructure, which is required for sperm motility9.
www.nature.com/reviews/molcellbio
870 | novEMBER 2007 | voluME 8

2007 Nature Publishing Group
REVIEWS
Box 1 | What happens to mitochondrial components after fusion?
Wild type
Fusion mutant
In time-lapse movies of labelled mitochondria in living cells, mitochondria are observed to undergo cycles of fusion and fission. With each fusion event, two mitochondria are Nature Reviews | Molecular Cell Biology merged into one. Intuitively, true fusion would be expected to involve outer membrane and inner membrane fusion, which would also result in mixing of the matrix contents. Indeed, these expectations have been experimentally confirmed. Apparent fusion events that have been visualized in cells can be confirmed by a cell-hybrid mitochondrial fusion assay6,32 (see figure). In this assay, mitochondria in two distinct cell lines are differentially labelled with mitochondrially targeted green fluorescent protein (GFP) and DsRed. The cell lines are co-plated onto cover slips and exposed briefly to polyethylene glycol, a chemical that induces adjacent cells to fuse into cell hybrids. After a recovery period, the cell hybrids are examined for mitochondrial fusion. In cell hybrids from normal cells, mitochondrial fusion results in mitochondria that carry both GFP and DsRed (see figure, top). Cells that are defective for mitochondrial fusion form cell hybrids with distinct red and green mitochondria (see figure, bottom). A conceptually similar assay can be performed with yeast cells by allowing labelled yeast strains to mate and form zygotes4. By using matrix-targeted fluorophores, these assays show that mitochondrial fusion results in mixing of the matrix contents. Moreover, by using mitochondrial markers that are localized to the outer or inner membranes, fusion of the individual membranes can be experimentally demonstrated; under normal conditions, outer and inner membrane fusion appear to be closely synchronized. An important question is what happens to mitochondrial DNA (mtDNA) after fusion. Each mitochondrion contains multiple copies of the mtDNA genome that are organized into one or more nucleoids. After fusion, these nucleoids appear to be motile and can potentially interact with each other73. In mammalian cells, mtDNA recombination has been documented, but its extent and importance is unclear.
lines of evidence from neuronal studies suggest that mitochondrial transport is regulated. First, mitochondria are recruited to regions with high energy demands, including active growth cones, presynaptic sites and postsynaptic sites13,16,17. Such recruitment is regulated by neuronal activation, further arguing that the recruitment of mitochondria is responsive to the local metabolic state. Second, neuronal mitochondria pause most often at sites that lack other mitochondria, resulting in a well-spaced axonal mitochondrial distribution14. Third, studies with the membrane-potential indicator dye JC-1 suggest that mitochondria with high membrane potential preferentially migrate in the anterograde direction, whereas mitochondria with low membrane potential move in the retrograde direction14. These migration patterns suggest that active mitochondria are recruited to distal regions with high energy requirements, whereas impaired mitochondria are returned to the cell soma, perhaps for destruction or repair. Finally, mitochondrial transport along axons responds to local concentrations of nerve growth factor (nGF), suggesting that specific signalling pathways control mitochondrial recruitment and retention16,18. Dynamic internal structure. In addition to changes in the overall shape of mitochondria, the internal structures of mitochondria are also dynamic. Three-dimensional tomography of cryopreserved samples has provided new views of inner membrane organization and plasticity19. The inner membrane can be divided into distinct regions: the inner boundary membrane, the cristae membrane and the cristae junctions (FIG.1c). The inner boundary membrane comprises the regions in which the inner membrane is in close proximity to the outer membrane. These regions are probably important for protein import and might be the sites of coupled outer and inner membrane fusion. The cristae junctions are narrow neck regions that separate the inner boundary membrane from the involuted cristae membrane. Cytochrome c, an intermembrane-space protein, is enriched in the space that is encased by cristae membranes, and the regulated opening of cristae junctions might be important for its relocalization during apoptosis20. These regions of the mitochondrial inner membrane are not only morphologically distinct but also appear to constitute separate functional domains. Proteins that are involved in the translocation of proteins through the inner membrane, such as the TIM23 complex, are enriched in the inner boundary membrane, whereas proteins that are involved in oxidativephosphorylation are enriched in the cristae membranes2123. In addition, the structure of mitochondrial membranes is linked to the metabolic state of mitochondria (FIG.1c). Purified mitochondria placed in low ADP conditions have limited respiration and have an orthodox morphology, characterized by narrow cristae and few cristae junctions per cristae compartment. under high ADP and substrate conditions, isolated mitochondria have high respiratory activity and a condensed morphology, characterized by larger cristae and several cristae junctions per cristae compartment19. It is unknown how inner membranes convert between these states, but inner membrane fusion
voluME 8 | novEMBER 2007 | 871
Anterograde
Thedirectionfromthecell bodytowardstheperiphery.
Retrograde
Thedirectionfromperipheral regionstowardsthecellbody.
Oxidative phosphorylation
AbiochemicalpathwayforATP productionthatresultsin oxygenconsumptionandis localizedtothemitochondrial cristae.
Dynamic subcellular distribution. Mitochondrial transport is required to distribute mitochondria throughout the cell (FIG.1b). In most cells, mitochondria are highly motile and travel along cytoskeletal tracks. Mitochondrial transport depends on the actin cytoskeleton in budding yeast10 and on both actin and microtubules in mammalian cells3,11,12. Depending on the cellular context, these transport processes can ensure proper inheritance of mitochondria or can recruit mitochondria to active regions of the cell. For example, in budding yeast, mitochondria are transported into and retained in the developing bud to ensure mitochondrial inheritance to the daughter cell10. This regulation of mitochondrial distribution is particularly evident in neurons. Quantitative measurements of neuronal mitochondrial transport have reported rates ranging from 0.4 mm min1 (ReF.13) to 0.11 mm sec1 (ReFs11,14,15). Such directed movements are not continuous; rather, they are saltatory, with pauses often followed by a reversal of direction. These patterns might reflect the attachment and detachment of cytoskeletal motors. Although these movements can appear chaotic, several
nATuRE REvIEWS | molecular cell biology

REVIEWS
a
Fusion + Fission
b
Anterograde movement Nucleus
member of the mitofusin family of GTPases. The yeast orthologue, Fzo1, has a conserved role in mitochondrial fusion24, and genetic screens in yeast have identified additional modulators of mitochondrial fusion and fission2,25 (FIG.3b). The core machineries that mediate mitochondrial fusion and fission are best understood in yeast. Several of these components have functionally conserved mammalian homologues. More comprehensive discussions of the molecular mechanisms of mitochondrial fusion and fission have been presented in recent reviews (for example, see ReF.1). Mitochondrial fusion. In yeast, the core mitochondrial fusion machinery consists of two GTPases: Fzo1 and Mgm1 (FIG.3). Fzo1 is located on the mitochondrial outer membrane and is essential for fusion of the outer membranes24,26. The mammalian homologues of Fzo1 are the mitofusins MFn1 and MFn2. These two related proteins form homo-oligomeric and hetero-oligomeric complexes that are essential for fusion6,27,28. Mitofusins are required on adjacent mitochondria during the fusion process, implying that they form complexes in trans between apposing mitochondria26,29. A heptad repeat region of MFn1 has been shown to form an antiparallel coiled coil that is probably involved in tethering mitochondria during fusion29. Mgm1 is a dynamin-related protein that is essential for fusion of the mitochondrial inner membranes in yeast30, a function that is consistent with its localization to the intermembrane space and its association with the inner membrane. The mammalian orthologue oPA1 is also essential for mitochondrial fusion28,31. In yeast, the outer membrane protein ugo1 physically links Fzo1 and Mgm1, but no mammalian orthologue has yet been discovered2. The membranepotential across the mitochondrial inner membrane is maintained by the electron transport chain and is essential for mitochondrial fusion26,32. Ionophores that dissipate the mitochondrial membrane potential cause mitochondrial fragmentation, owing to an inhibition of mitochondrial fusion32,33. In an in vitro fusion assay, both the proton and the electrical gradient components of the membrane potential are important26. The mechanistic link between membrane potential and fusion remains to be resolved, but one factor appears to be the dependence of post-translational processing of oPA1 on the membrane potential34. Recent work has also identified mitochondrial lipids as important factors in fusion. Mitochondrial morphology screens in yeast identified members of the ergosterol synthesis pathway as being required for normal mitochondrial morphology35,36. Recently, mitochondrial phospholipase D has been identified as a protein that is important for mitochondrial fusion37. This mitochondrial outer membrane enzyme hydrolyses cardiolipin to generate phosphatidic acid. Interestingly, ergosterol has been associated with yeast vacuole fusion38, and phosphatidic acid is thought to play a part in generating the membrane curvature that is required for sNARe-mediated fusion39. Thus, specific lipids might have similar roles in distinct types of membrane fusion.
Retrograde movement
c
IMS CJ IBM CM Condensed morphology Orthodox morphology OM IM
Low [ADP] High [ADP]
Figure 1 | mitochondria as dynamic organelles. a | Mitochondrial fusion and fission control mitochondrial number and size. With fusion, Nature Reviews | Molecular Cell Biology two mitochondria become a single larger mitochondrion with continuous outer and inner membranes. Conversely, a single mitochondrion can divide into two distinct mitochondria by fission. b | In mammalian systems, mitochondria are distributed throughout the cytoplasm by active transport along microtubules and actin filaments. Distinct molecular motors transport the mitochondria in anterograde or retrograde directions. c | Inner membrane dynamics. The diagram indicates the different regions of the inner membrane. The bottom panels show electron microscopy (EM) tomograms of two mitochondria under different metabolic conditions (red, outer membrane; yellow, inner boundary membrane; green, cristae membrane). Cristae organization can vary widely, often in response to the bioenergetic state of the cell: an orthodox cristae morphology, with narrow cristae and few cristae junctions per cristae compartment, is found in low ADP conditions, whereas a condensed morphology, with larger cristae and several junctions per cristae compartment, is found in high ADP conditions19. EM images reproduced with permission from ReF.19 (2006) Elsevier. CJ, cristae junction; CM, cristae membrane; IBM, inner boundary membrane; IM, inner membrane; IMS, intermembrane space; OM, outer membrane.
Coiled coil
Astructuralmotifthatis formedbypolypeptide sequencesthatcontain hydrophobicheptadrepeats.
and fission might be involved19. Taken together, these observations indicate that inner membrane morphology is intimately related to bioenergetics, although the causal relationship remains unclear.
Dynamin
AlargeGTPasethatisthought tomediatevesiclescission duringendocytosis.
Mediators of fusion and fission Molecular analysis of mitochondrial morphology began with the discovery in 1997 of the D. melanogaster fusion factor fuzzy onions (FZo), a mitochondrial outer membrane GTPase that is required for the fusion of mitochondria during spermatogenesis9. FZo is the founding

REVIEWS
No fusion Mfn-null Wild type No fission DRP1 K38A
Figure 2 | mitochondrial fusion and fission regulate morphology. Mitochondrial length, size and connectivity are determined by theNature Reviews | Molecular Cell Biology relative rates of mitochondrial fusion and fission. In wild-type cells (shown in the central panel), mitochondria form tubules of variable length. In the absence of mitochondrial fusion (for example, in mitofusin (Mfn)null cells (shown in the left panel), which lack MFN1 and MFN2), unopposed fission results in a population of mitochondria that are all fragmented. Conversely, decreased fission relative to fusion (for example, in DRP1 K38A cells (shown in the right panel), which have a dominant-negative form of dynamin-related protein-1 (DRP1)) results in elongated and highly interconnected mitochondria. Scale bar represents 10 mm.
Mitochondrial membrane potential

Theelectrochemicalgradient thatexistsacrossthe mitochondrialinner membrane.
Ergosterol
Asteroidcompoundthatisa componentofyeastcell membranesandwhichmight havearolesimilartothatof cholesterolinmammaliancell membranes.
SNARE
(solubleNethylmaleimide sensitivefusionprotein(NsF) attachmentprotein(sNAP) receptor).Ahighlyhelical proteinthatmediatesthe specificfusionofvesicleswith targetmembranes.
F-box protein
AproteincontaininganFbox motif,asmalldomainthatis usedforproteininteractions. ThebestcharacterizedFbox proteinsarecomponentsofan e3ubiquitinligase,andhelpin ubiquitindependentprotein degradationbyrecognizing specificsubstrates.
Mitochondrial fission. Mitochondrial fission requires the recruitment of a dynamin-related protein (Dnm1 in yeast and DRP1 in mammals) from the cytosol (FIG.4). Both Dnm1 and DRP1 assemble into punctate spots on mitochondrial tubules, and a subset of these complexes lead to productive fission events5,7,8. By analogy with the classical function of dynamin in endocytosis, Dnm1 and DRP1 are thought to assemble into rings and spirals that encircle and constrict the mitochondrial tubule during fission25. Consistent with this model, purified Dnm1 can indeed form helical rings and spirals in vitro, with dimensions that are similar to those of constricted mitochondria40. Moreover, Dnm1 assembly is required for fission activity41. The recruitment of Dnm1 to yeast mitochondrial fission sites involves three other components. one of these is Fis1, a mitochondrial integral outer membrane protein that is essential for fission4244. Fis1 binds indirectly to Dnm1 through one of two molecular adaptors, Mdv1 or Caf4 (ReF.45) (FIG.4b). Either Mdv1 or Caf4 is sufficient to allow the Fis1-dependent recruitment of Dnm1, although Mdv1 has a more important role in mediating fission. FIS1, the mammalian homologue of Fis1, is also essential for mitochondrial fission46, but no homologues of Mdv1 or Caf4 are currently known. FIS1 and DRP1 are also required for the fission of peroxisomes47,48.
This modification of DRP1 might affect its association with mitochondrial membranes. Mitochondrial fission is also regulated by the cell cycle. For example, mitochondria in Hela cells are usually tubular, but they become more fragmented during mitosis, a phenomenon that might facilitate the partitioning of mitochondria to daughter cells during cytokinesis. This regulated fragmentation of mitochondria is due to increased mitochondrial fission, and phosphorylation of DRP1 during mitosis has been implicated55. In addition to the genes that encode core fusion and fission components, other genes can affect mitochondrial morphology. large-scale visual screens for aberrant mitochondrial morphology in mutant yeast have yielded numerous genes of interest and provided general insights into the control of mitochondrial morphology35,36. These screens suggest that several cellular pathways influence mitochondrial morphology and inheritance, including ergosterol biosynthesis, mitochondrial protein import, actin dynamics, vesicular fusion and ubiquitin-mediated protein degradation. The close interplay between mitochondrial protein import and morphology has been emphasized by the recent finding that the mitochondrial morphology genes MMM1, MDM10 and MDM12 have a direct role in the assembly of barrelproteins in the outer mitochondrial membrane56. Proteins that are required for mitochondrial transport. Energy-dependent molecular motors transport mitochondria along cytoskeletal filaments. Along microtubules, multiple kinesin family members and cytoplasmic dynein have been implicated in anterograde and retrograde mitochondrial transport, respectively3. Recent work has clarified the linkage between mitochondria and kinesin-1. Genetic screens in D. melanogaster identified milton and Miro, both of which are required for anterograde mitochondrial transport in neurons57,58. Milton interacts directly with kinesin and Miro, which is a mitochondrial outer membrane protein that has GTPase and Ca2+-binding eFhanddomains59. In yeast, disruption of the Miro orthologue Gem1 results in abnormalities in mitochondrial morphology and poor respiratory activity60. Both GTP-binding and Ca2+-binding motifs are essential for Gem1 function, which appears not to be involved in fusion or fission. Depending on the cell type, mitochondria can also travel along actin filaments under the control of myosin motors3. Proteins that mediate inner membrane morphology. Studies of mitochondrial inner membrane structure are complicated by the intimate link between mitochondrial bioenergetics and cristae structure. As a result, disruption of the proteins that are important for bioenergetics can lead to a secondary effect on inner membrane structure. nevertheless, several proteins probably have a specific role in controlling cristae structure. In addition to their roles in mitochondrial fusion, Mgm1 and oPA1 are important for cristae structure. loss of Mgm1 in yeast or knockdown of oPA1 in mammalian cells results in disorganized inner membrane structures30,6164. In both cases, homo-oligomeric interactions are involved30,64.
-barrel protein
Aproteincomposedofa sheetthatisrolledup intoacylinder.Onesuch mitochondrialbarrelprotein isVDAC(voltagedependent anionchannel),whichformsa poreintheoutermembrane.
Kinesin
Amicrotubulebasedmolecular motorproteinthatismost oftendirectedtowardstheplus endofmicrotubules.
Other regulators of dynamics Mitochondrial fusion and fission activities are probably coordinated with cellular physiology. In yeast, the steadystate levels of Fzo1 are controlled by the Fboxprotein Mdm30, which negatively regulates Fzo1 levels in a proteasome-independent manner49,50. In mammalian cells, post-translational modification of DRP1 regulates its function in mitochondrial fission. The mitochondrial E3 ubiquitin ligase MARCH5 is essential for mitochondrial fission51. This requirement is probably related to the ability of MARCH5 to promote DRP1 ubiquitylation and to associate physically with ubiquitylated DRP1 (ReFs52,53). Furthermore, during apoptosis, sumoylation of DRP1 is activated in a BAX- and/or BAK-dependent manner54.

REVIEWS
a
OM fusion IM fusion
Fzo1
mitofilin in mammalian cells causes dramatic abnormalities of the cristae, resulting in the formation of complex layers of inner membrane69. Depletion of Mmm1, Mdm31 and Mdm32 yeast proteins implicated in mitochondrial(mt)DNA maintenance also cause aberrant cristae morphologies70,71.
Ugo1 s-Mgm1 I-Mgm1 IMS OM
IM
Cristae
Dynein
Amicrotubulebasedmolecular motorthatisdirectedtowards theminusendofmicrotubules.
EF-hand domain
Ahelixloophelixproteinmotif thatcanbindaCa2+ion.
Figure 3 | mitochondrial fusion. a | Mitochondrial fusion consists of outer membrane (OM) fusion followed by inner membrane (IM) fusion. Normally these events occur Nature Reviews | Molecular Cell Biology coordinately. b | The dynamin-related proteins Fzo1 and Mgm1 are key molecules in the yeast mitochondrial fusion machinery. Fzo1 is an integral outer membrane protein with GTPase and heptad repeat domains that face the cytoplasm. All of the domains are required for the fusion activity of Fzo1. Mgm1 is present on the inner membrane, facing the intermembrane space (IMS), and is proteolytically processed by a rhomboid protease. Both long (l-Mgm1) and short (s-Mgm1) forms are required for mitochondrial fusion. In addition to inner membrane fusion, Mgm1 is required for the maintenance of cristae structures. Ugo1 binds to both Fzo1 and Mgm1 and probably coordinates their function. All components are encoded by nuclear DNA. The mitofusin proteins MFN1 and MFN2 are the mammalian homologues of Fzo1; OPA1 is the mammalian homologue of Mgm1. No mammalian homologue of Ugo1 has been identified so far.
Biological functions of mitochondrial dynamics Why do mitochondria continually fuse and divide? Recent studies show that these processes have important consequences for the morphology, function and distribution of mitochondria. First, fusion and fission control the shape, length and number of mitochondria. The balance between these opposing processes regulates mitochondrial morphology. Second, fusion and fission allow mitochondria to exchange lipid membranes and intramitochondrial content. Such exchange is crucial for maintaining the health of a mitochondrial population. Third, the shape of mitochondria affects the ability of cells to distribute their mitochondria to specific subcellular locations. This function is especially important in highly polarized cells, such as neurons. Finally, mitochondrial fission facilitates apoptosis by regulating the release of intermembrane-space proteins into the cytosol. As a result of these cellular functions, mitochondrial dynamics has consequences for development, disease and apoptosis.
Maintaining a healthy mitochondrial population. Mitochondrial fusion is required to maintain a functional mitochondrial population in the cell. Fibroblasts that lack both MFn1 and MFn2 have reduced respiratory capacity, and individual mitochondria show great heterogeneity in shape and membrane potential28. Cells that lack oPA1 show similar defects, with an even greater reduction in respiratory capacity. How does fusion protect mitochondrial function? It is probable that a primary function of mitochondrial fusion is to enable the exchange of contents between mitochondria (BOX1). As a result, mitochondria should not be considered autonomous organelles; instead, the hundreds of mitochondria in a typical cell exist as a population of organelles that cooperate with each other through fusion and fission. The heterogeneous properties of mitochondria in fusion-deficient cells are consistent with this model28. In normal cells, a few mitochondria might be non-functional owing to the loss of essential components. However, this dysfunction is transient because mitochondrial fusion provides a pathway for these defective mitochondria to regain essential components (FIG.5a). An essential component of mitochondrial function is mitochondrial DnA (mtDnA), which is organized into compact particles termed nucleoids. The mtDnA genome encodes essential subunits of the respiratory complexes I, III and Iv, and is therefore essential for oxidative phosphorylation. When mitochondrial fusion is abolished, a large fraction of the mitochondrial population loses mtDnA nucleoids72. During mitochondrial division in normal cells, most daughter mitochondria inherit at least one mtDnA nucleoid73. However, in cases where a
Mitochondrial F1F0 ATP synthase

Alarge,multisubunitenzyme embeddedinthe mitochondrialcristaethatuses theprotongradientacrossthe innermembranetosynthesize ATP.
Mitochondrial DNA
(mtDNA).Acirculargenome (~16kbinmammals)located inthemitochondrialmatrix thatencodes13polypeptides oftheelectrontransportchain, 22tRNAsand2rRNAs.
Nucleoid
AcompactedmassofDNA. MitochondrialDNAis organizedintonucleoids,each consistingofseveral mitochondrialgenomes.
Mitochondrial F 1FoATP synthase , a rotary enzyme embedded in the inner membrane that couples proton pumping to ATP synthesis, is essential for normal cristae structure65. This role in inner membrane structure involves a dimeric form of ATP synthase that contains the additional subunits e and g. As visualized by electron microscopy, the ATP synthase dimer has a dimeric interface with a sharp angle that could distort the local lipid membrane. This distortion might contribute to the high membrane curvature that characterizes cristae tubules66,67. Mgm1 is required for the oligomerization of ATP synthase, providing a link between these two modulators of cristae structure63. Additional proteins modulate inner membrane dynamics. In yeast, Mdm33 is required for normal mitochondrial morphology and its overexpression leads to septation and vesiculation of the inner membranes68. Because of these phenotypes, Mdm33 has been suggested to have a role in inner membrane fission. Knockdown of

REVIEWS
a
Dnm1
b
Dnm1
Adaptor poteins (Mdv1, Caf4)
Fis1 OM IM
Figure 4 | mitochondrial fission. a | In yeast, mitochondrial fission is mediated by the dynamin-related protein Dnm1. Cytoplasmic Dnm1Nature Reviews | mitochondrial outer localizes to the Molecular Cell Biology membrane (OM), where it oligomerizes into a ring structure that constricts and severs the mitochondrion. In this model, Dnm1 functions in an analogous manner to the way dynamin functions in endocytosis. b | The localization of Dnm1 on the mitochondrial outer membrane is mediated by Fis1 and the adaptor proteins Mdv1 and Caf4. Fis1 is an integral outer membrane protein that interacts with the N termini of Mdv1 and Caf4. Both Mdv1 and Caf4 have C-terminal WD-40 repeats that bind Dnm1. Fis1 and Dnm1 have mammalian homologues (FIS1 and DRP1, respectively), but no Mdv1 or Caf4 homologues have been identified so far. IM, inner membrane.
daughter fails to inherit a nucleoid, mitochondrial fusion would enable restoration of mtDnA. In fusion-deficient cells, the lack of content exchange prevents restoration of mtDnA nucleoids and probably accounts for the heterogeneity in membrane potential and the reduced respiratory capacity. It should be noted that fusiondeficient cells still maintain significant numbers of mtDnA nucleoids; however, due to ongoing mitochondrial fission, these nucleoids are encased by a small mitochondrial mass, and therefore the functional mitochondrial mass (at least in terms of bioenergetics) in such cells is greatly reduced (FIG.5b). In addition to mtDnA, it is also possible that other components, such as substrates, metabolites or specific lipids, can be restored in defective mitochondria by fusion. Further studies will determine whether content exchange is the primary function of mitochondrial fusion. The importance of mitochondrial fusion in development and disease might be a consequence of this function. Essential developmental functions. Perturbations in mitochondrial dynamics result in specific developmental defects. Mice that lack either MFn1, MFn2 or oPA1 fail to survive past mid-gestation6,74,75. MFn2 has a highly specific function in the development of the trophoblast giant cell layer of the placenta6. likewise, MFn1 appears to have an essential placental function72. Mitochondrial fission is also an essential process. Worms that are deficient in mitochondrial division die before adulthood76. An infant patient with a dominantnegative DRP1 allele has been reported. This patient died at ~1 month of age and had a wide range of abnormalities, including reduced head growth, increased lactic acid and optic atrophy. Fibroblasts from this patient showed elongated mitochondria and peroxisomes 77. It is unclear how the developmental defects are related to these organellar shape changes.
Mitochondrial distribution and recruitment in neurons. Given the importance of mitochondrial dynamics in maintaining bioenergetics, these dynamics are probably a ubiquitous phenomenon that is important for all cells. However, certain cells, particularly neurons, seem to be especially dependent on its proper control. This dependence of neurons probably stems from their high energy demands and the special importance of proper mitochondrial distribution: mitochondria are concentrated in several neuronal regions, including pre- and postsynaptic sites13,17. To achieve this non-uniform distribution, neurons rely heavily on active transport to recruit mitochondria and other organelles to nerve terminals3. The proper localization of mitochondria to axon terminals depends on mitochondrial dynamics. neurons that lack Milton, Miro or DRP1 show defective mitochondrial transport and have sparse mitochondria at axon terminals. Such distribution defects lead to reduced capacity for synaptic transmission57,58,78. It seems likely that mitochondria that are localized to synapses are primarily required to drive ATP-dependent processes. The synapses of neurons that express mutant DRP1 show defective mobilization of the reserve vesicle pool (an ATPdependent process), and the defects in synaptic transmission can be rescued by experimentally filling synapses with ATP. In addition, synapse-localized mitochondria help to regulate Ca2+ homeostasis, although this function appears to be crucial only during intense synaptic activity. Synapses that lack Miro or DRP1 have elevated resting Ca2+ levels, but normal Ca2+ dynamics are maintained except under sustained nerve stimulation57,78. Both mitochondrial fusion and fission affect the mitochondrial distribution in dendrites. In hippocampal neurons, mitochondria accumulate at dendritic spines following neuronal stimulation13. Inhibition of mitochondrial fission causes elongation of the mitochondria and decreases the abundance of dendritic mitochondria and the density of dendritic spines. Conversely, increased fission facilitates the mobilization of dendritic mitochondria and leads to an increased spine number13. In the cerebellum, the distribution of mitochondria in the dendritic processes of Purkinje neurons is highly dependent on mitochondrial fusion72 (see below). Lymphocyte chemotaxis. Mitochondrial dynamics appears to be important for proper mitochondrial redistribution in lymphocytes during chemotaxis 79. Mitochondria are concentrated in the trailing edge in lymphocyte cell lines that migrate in response to chemical attractants. Modulation of mitochondrial fusion or fission affects both mitochondrial redistribution and cell migration. Fragmentation enhances mitochondrial redistribution and cell migration, whereas conditions that promote fusion have the opposite effect. Therefore, as in neurons, mitochondrial shape in lymphocytes can affect the recruitment of mitochondria to local cellular areas. Regulation of apoptosis. In apoptosis, several structural changes occur in mitochondria during the early phase of cell death (FIG.6). The mitochondria become fragmented owing to increased fission activity. At approximately the
Chemotaxis
Thedirectedmovementofcells inresponsetoachemical stimulus.

REVIEWS
a Wild-type cells b Fusion-deficient cells
Fusion
Defective mitochondrion
Fission
Rescued mitochondrion
Figure 5 | mitochondrial dynamics protects mitochondrial function. a | In wild-type Nature Reviews | Molecular Cell Biology cells, the vast majority of mitochondria are functional (shown in green). In this simplified diagram, one mitochondrion is depicted as non-functional (shown in orange). One of several possible reasons for dysfunction is a lack of mitochondrial DNA (mtDNA) nucleoids (shown as black circles). The dysfunctional mitochondrion can regain its function and mtDNA by fusing with a neighbouring mitochondrion. The fused mitochondrion then undergoes fission, with both daughter mitochondria receiving mtDNA nucleoids. It should be noted that the identities of the daughter mitochondria are distinct from the parental mitochondria, owing to content exchange and the fact that the fission point is typically distinct from the fusion point. b | In fusion-deficient cells, mitochondria are fragmented due to ongoing fission in the absence of fusion. Mitochondria that lack mtDNA nucleoids accumulate because there is no pathway for defective mitochondria to regain mtDNA. Fusion-deficient cells can maintain mtDNA nucleoids, but such nucleoids serve a much smaller mitochondrial mass.
can occur in the absence of mitochondrial fission. An important issue to resolve in future studies is how fission is related to the permeabilization of mitochondria. Surprisingly, the apoptotic proteins BAX and BAK, which have well-established pro-apoptotic roles in mitochondrial membrane permeabilization, also appear to regulate mitochondrial morphology. BAX and BAK double-knockout cells have fragmented mitochondria due to reduced mitochondrial fusion87, although the extent of this effect depends on the experimental system88. little is known about how BAX and BAK mediate their effects on mitochondrial morphology, but BAX influences MFn2 distribution on the mitochondrial outer membrane87 and BAK associates with MFn1 and MFn2 (ReF.88). In conjunction with MoMP, remodelling of the cristae membranes is required for the rapid and efficient release of cytochrome c20,89. Most cytochrome c is localized to cristae compartments20; oPA1 appears to regulate the diameter of cristae junctions and therefore regulates cytochrome c release 64,90. overexpression of oPA1 blocks cytochrome c release following the induction of apoptosis by maintaining narrow cristae junctions64. DRP1 has also been proposed to play a part in cristae remodelling during apoptosis91.
Role in human disease Several human diseases are caused by mutations in genes that are essential for mitochondrial dynamics (TABLe1). Each of these diseases causes degeneration of specific nerves, reinforcing the notion that neurons are particularly prone to defects in mitochondrial dynamics.
OPA1 and autosomal dominant optic atrophy. Heterozygous mutations in oPA1 cause autosomal dominant optic atrophy (ADoA), the most common heritable form of optic neuropathy92,93. This disease is characterized by the degeneration of retinal ganglion cells, the axons of which form the optic nerve. More than 100 pathogenic oPA1 mutations have been reported, with most occurring in the GTPase domain94. Half of the mutants encode a truncated protein owing to a nonsense mutation. A few nonsense mutations abolish nearly the entire coding sequence, suggesting that haploinsufficiency of oPA1 can cause ADoA. It remains possible that other, less severe, truncations might have dominant-negative activity. How these oPA1 mutations cause the clinical symptoms of ADoA remains to be clarified. non-neuronal cells from patients with ADoA can have aggregated, fragmented or normal mitochondria93,95; however, because data from only a few patients have been reported, it is not clear whether these findings are the norm. In addition, oPA1 mutations have been associated with reduced ATP production and reduced mtDnA content96,97. The defects that have been documented in human ADoA diseased tissue are not as severe as those observed in experimental cells in which oPA1 is depleted. Fibroblasts that are deficient for oPA1 have fragmented mitochondria, defects in respiration, aberrant cristae structure and increased susceptibility to apoptosis28,31,62,98.
Mitochondrial outer membrane permeabilization

(MOMP).Theopeningofpores inthemitochondrialouter membraneanearlyevent duringapoptosisthatreleases apoptoticfactorsfromthe mitochondrialintermembrane space.
Haploinsufficiency
Ageneticstateindiploidsin whichasinglefunctionalcopy ofageneisinsufficientto maintainanormalphenotype.
same time, mitochondrialoutermembranepermeabilization (MoMP) causes the release of contents of the intermembrane space, such as cytochrome c and second mitochondria-derived activator of caspase (SMAC)/ Diablo, into the cytoplasm. Because cytochrome c is preferentially sequestered in cristae compartments, it is thought that the opening of cristae junctions is a vital step in facilitating its efficient release. once in the cytosol, cytochrome c activates a cascade of caspases that propagate and execute the apoptotic programme. These three structural changes fragmentation, MoMP and cristae remodelling occur at similar times, but their temporal sequence and causative links are still controversial80,81. Mitochondrial fragmentation during apoptosis is associated with dynamic changes in the mitochondrial localization of several proteins, including BAX, BAK, MFn2, endophilin and DRP1 (ReF.81). Inhibition of fission activity blocks mitochondrial fragmentation, reduces cytochrome c release and can reduce or delay cell death depending on the experimental system46,82,83. In Caenorhabditis elegans and D. melanogaster, disruption of DRP1 reduces the number of cell deaths8486. In multiple systems, it seems that fission is important for rapid and efficient cell death, although apoptosis

REVIEWS
Cytochrome c
Apoptotic stimuli
Cristae junctions
Figure 6 | mitochondrial dynamics during apoptosis. At an early stage of apoptosis, Nature Reviews | Molecular Cell Biology three structural changes occur in mitochondria. Fragmentation takes place as a result of increased fission mediated by dynamin-related protein-1 (DRP1) and the mitochondrial fission-1 protein (FIS1). Mitochondrial outer membrane permeabilization (MOMP; indicated by dashed outlines) is induced by the pro-apoptotic BCL2-family members BAX and BAK. MOMP enables the release of cytochrome c (shown as red dots) and other soluble proteins from the intermembrane space. However, release of cytochrome c is efficient only if the cristae junctions are widened to allow escape from the cristae compartments. The dynamin-related proteins OPA1 and DRP1 have been implicated in cristae remodelling.
Mouse models of ADoA that contain oPA1 mutations develop the features of ADoA in an age-dependent manner74,75. Heterozygous mice show a progressive decline in retinal ganglion cell number and aberrations of axons in the optic nerve. Mice that are homozygous for the oPA1 mutation die at mid-gestation74,75, which is consistent with an essential requirement for mitochondrial fusion during embryonic development6. MFN2 and Charcot-Marie-Tooth 2A. Charcot-MarieTooth (CMT) disease, one of the most common hereditary neuropathies, is caused by mutations in at least 30 different genes99. Affected individuals have progressive distal motor and sensory impairments that start in the feet and hands as a result of the degeneration of the long peripheral nerves. Depending on the type of CMT, these diseases are caused by either a primary defect in the Schwann cells that myelinate the peripheral nerves or by a defect in the neurons themselves99. CMT2A is an axonopathy that is caused by the latter type of defect, and it has been associated with >40 mutations in MFn2. nearly all of these disease alleles contain missense mutations or short, in-frame deletions100. Most mutations cluster in or near the GTPase domain, but some also
Sural nerve
Asensorynerveinnervating thecalfandfootthatis commonlyinvestigatedby biopsyfortheevaluationof peripheralneuropathies.
Table 1 | Disorders associated with mitochondrial perturbations

Disease
CMT2A ADOA CMT4A Unnamed
mitochondrial function
Fusion Fusion Fission? Fission
gene
MFN2 OPA1 GDAP1 DRP1
Description
Autosomal dominant peripheral neuropathy Autosomal dominant optic atrophy (ADOA) Autosomal recessive peripheral neuropathy Neonatal lethality
CMT, Charcot-Marie-Tooth; DRP1, dynamin-related protein-1; GDAP1, ganglioside-induced differentiation-associated protein-1; MFN2, mitofusin-2; OPA1, optic atrophy-1.
occur in each of the heptad repeat domains of MFn2. In addition to the loss of peripheral nerve function, a subset of patients with CMT2A have optic atrophy, suggesting that oPA1 and MFn2 mutations can lead to overlapping clinical outcomes101,102. Because of the difficulties in studying nerve tissue from patients, the pathogenic mechanisms that lead to peripheral nerve degeneration in CMT2A are not well understood. only one study has reported ultrastructural defects in mitochondria from the nerves of patients with CMT2A. Mitochondria in the suralnerve of two patients showed structural aberrations in their outer and inner membranes, along with swelling that is suggestive of mitochondrial dysfunction103. Aggregation of mitochondria was also observed. Interestingly, CMT2A alleles of MFN2 (ReFs27,104) cause mitochondrial aggregation and subsequent mitochondrial transport defects in neurons104. However, the mitochondrial aggregation phenotype is dependent on significant overexpression27, and therefore its relevance to disease pathogenesis remains to be clarified. Several perplexing issues remain to be resolved concerning the molecular genetics of CMT2A. How does mutation of one copy of MFN2 lead to disease? Why are long peripheral neurons selectively affected, given that MFn2 is a broadly expressed protein? Clues to these issues have come from analysis of CMT2A alleles in mice27. Many CMT2A alleles of Mfn2 are non-functional for fusion when expressed alone. However, the fusion activity of these non-functional alleles can be efficiently complemented by wild-type MFn1 (but not MFn2). This complementation is due to the ability of MFn1 and MFn2 to form hetero-oligomeric complexes that are functional for fusion. In a patient with CMT2A, therefore, cells that express MFn1 are protected from gross loss of fusion activity. By contrast, cells with little or no MFn1 expression suffer a greater relative loss of fusion activity. In part, these properties of the CMT2A alleles might underlie the selective loss of sensory and motor neurons. Consistent with this model, MFn2 seems to be more highly expressed in central and peripheral nervous tissue than MFn1 (S.A.D. and D.C.C., unpublished observations). Even in the peripheral nerves, it appears that mitochondrial fusion defects are only partial because only the longest nerves are affected. Most probably, the extreme dimensions of the long peripheral nerves make them most vulnerable to changes in mitochondrial dynamics. How might perturbations in mitochondrial fusion lead to neurodegeneration? Clues to the pathogenic mechanisms have come from the finding that mice that lack MFn2 show highly specific degeneration of Purkinje neurons in the cerebellum, resulting in cerebellar ataxia72. Purkinje cells are the sole efferent neurons of the cerebellum, and they have exquisitely formed dendritic processes. Both developing and mature Purkinje cells that lose MFn2 fail to support dendritic outgrowth, particularly that of dendritic spines, which are the sites of synaptic connections. In normal Purkinje cells, abundant tubular mitochondria reside in dendritic processes. By contrast, mutant Purkinje cells have fragmented mitochondria that fail to distribute effectively along dendritic

REVIEWS
processes. In addition, the Purkinje cells show a loss of respiratory activity, probably owing to an accumulation of mitochondria that lack mtDnA nucleoids. Therefore, loss of mitochondrial fusion in Purkinje neurons impairs respiratory activity and mitochondrial localization. GDAP1 and Charcot-Marie-Tooth 4A. Another form of CMT is associated with defects in mitochondrial dynamics. Ganglioside-induced differentiation-associated protein-1 (GDAP1) is mutated in CMT4A, one of the few recessive forms of CMT disease. CMT4A has both demyelinating and axonal features and, consistent with this mixed clinical presentation, GDAP1 is expressed in both Schwann cells and neurons105. GDAP1 is an integral outer membrane protein that probably affects mitochondrial division105. Disease alleles either fail to localize to mitochondria or are defective in stimulating mitochondrial fission when overexpressed105. If GDAP1 disease alleles disrupt normal mitochondrial fission, they might cause mitochondrial distribution defects similar to those that are induced by the DRP1 mutations discussed above13,78.
1. 2. 3. 4. Chan, D. C. Mitochondrial fusion and fission in mammals. Annu. Rev. Cell Dev. Biol. 22, 7999 (2006). Okamoto, K. & Shaw, J. M. Mitochondrial morphology and dynamics in yeast and multicellular eukaryotes. Annu. Rev. Genet. 39, 50336 (2005). Hollenbeck, P. J. & Saxton, W. M. The axonal transport of mitochondria. J. Cell Sci. 118, 54115419 (2005). Nunnari, J. et al. Mitochondrial transmission during mating in Saccharomyces cerevisiae is determined by mitochondrial fusion and fission and the intramitochondrial segregation of mitochondrial DNA. Mol. Biol. Cell 8, 12331242 (1997). Bleazard, W. et al. The dynamin-related GTPase Dnm1 regulates mitochondrial fission in yeast. Nature Cell Biol. 1, 298304 (1999). Chen, H. et al. Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J. Cell Biol. 160, 189200 (2003). Sesaki, H. & Jensen, R. E. Division versus fusion: Dnm1p and Fzo1p antagonistically regulate mitochondrial shape. J. Cell Biol. 147, 699706 (1999). Smirnova, E., Griparic, L., Shurland, D. L. & van der Bliek, A. M. Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol. Biol. Cell 12, 22452256 (2001). Hales, K. G. & Fuller, M. T. Developmentally regulated mitochondrial fusion mediated by a conserved, novel, predicted GTPase. Cell 90, 121129 (1997). This study identified the first component of the mitochondrial fusion machinery, thereby providing an avenue to identify new components in further yeast genetic screens. Fehrenbacher, K. L., Yang, H. C., Gay, A. C., Huckaba, T. M. & Pon, L. A. Live cell imaging of mitochondrial movement along actin cables in budding yeast. Curr. Biol. 14, 19962004 (2004). Morris, R. L. & Hollenbeck, P. J. Axonal transport of mitochondria along microtubules and F-actin in living vertebrate neurons. J. Cell Biol. 131, 13151326 (1995). Ligon, L. A. & Steward, O. Role of microtubules and actin filaments in the movement of mitochondria in the axons and dendrites of cultured hippocampal neurons. J. Comp. Neurol. 427, 351361 (2000). Li, Z., Okamoto, K., Hayashi, Y. & Sheng, M. The importance of dendritic mitochondria in the morphogenesis and plasticity of spines and synapses. Cell 119, 873887 (2004). Miller, K. E. & Sheetz, M. P. Axonal mitochondrial transport and potential are correlated. J. Cell Sci. 117, 27912804 (2004).
Perspectives The study of mitochondrial dynamics has undergone great advances in the past few years. It is now clear that mitochondrial dynamics is important for the functional state of mitochondria. By enabling content exchange between mitochondria, fusion and fission prevent the accumulation of defective mitochondria. These opposing processes also control mitochondrial shape, which affects the distribution of mitochondria as well as their participation in apoptosis. As a result, mitochondrial dynamics is particularly important in cells and tissues that have a special dependence on mitochondrial function. Defects in mitochondrial dynamics can manifest in mammalian development, apoptosis and disease. As our knowledge of mitochondrial dynamics increases, we can expect to learn about its involvement in other processes. The link between defects in mitochondrial fusion and neurodegenerative disease is particularly intriguing. In future studies, the pathophysiological mechanisms that underlie neurodegenerative diseases such as ADoA and CMT2A will hopefully be further dissected in appropriate animal models.
30. Meeusen, S. et al. Mitochondrial inner-membrane fusion and crista maintenance requires the dynaminrelated GTPase Mgm1. Cell 127, 383395 (2006). 31. Cipolat, S., Martins de Brito, O., Dal Zilio, B. & Scorrano, L. OPA1 requires mitofusin 1 to promote mitochondrial fusion. Proc. Natl Acad. Sci. USA 101, 1592715932 (2004). 32. Legros, F., Lombes, A., Frachon, P. & Rojo, M. Mitochondrial fusion in human cells is efficient, requires the inner membrane potential, and is mediated by mitofusins. Mol. Biol. Cell 13, 43434354 (2002). 33. Malka, F. et al. Separate fusion of outer and inner mitochondrial membranes. EMBO Rep. 6, 853859 (2005). 34. Ishihara, N., Fujita, Y., Oka, T. & Mihara, K. Regulation of mitochondrial morphology through proteolytic cleavage of OPA1. EMBO J. 25, 29662977 (2006). 35. Altmann, K. & Westermann, B. Role of essential genes in mitochondrial morphogenesis in Saccharomyces cerevisiae. Mol. Biol. Cell 16, 54105417 (2005). 36. Dimmer, K. S. et al. Genetic basis of mitochondrial function and morphology in Saccharomyces cerevisiae. Mol. Biol. Cell 13, 847853 (2002). 37. Choi, S. Y. et al. A common lipid links Mfn-mediated mitochondrial fusion and SNARE-regulated exocytosis. Nature Cell Biol. 8, 12551262 (2006). 38. Fratti, R. A., Jun, Y., Merz, A. J., Margolis, N. & Wickner, W. Interdependent assembly of specific regulatory lipids and membrane fusion proteins into the vertex ring domain of docked vacuoles. J. Cell Biol. 167, 10871098 (2004). 39. Vitale, N. et al. Phospholipase D1: a key factor for the exocytotic machinery in neuroendocrine cells. EMBO J. 20, 24242434 (2001). 40. Ingerman, E. et al. Dnm1 forms spirals that are structurally tailored to fit mitochondria. J. Cell Biol. 170, 10211027 (2005). 41. Bhar, D., Karren, M. A., Babst, M. & Shaw, J. M. Dimeric Dnm1-G385D interacts with Mdv1 on mitochondria and can be stimulated to assemble into fission complexes containing Mdv1 and Fis1. J. Biol. Chem. 281, 1731217320 (2006). 42. Fekkes, P., Shepard, K. A. & Yaffe, M. P. Gag3p, an outer membrane protein required for fission of mitochondrial tubules. J. Cell Biol. 151, 333340 (2000). 43. Mozdy, A. D., McCaffery, J. M. & Shaw, J. M. Dnm1p GTPase-mediated mitochondrial fission is a multi-step process requiring the novel integral membrane component Fis1p. J. Cell Biol. 151, 367380 (2000). 44. Tieu, Q. & Nunnari, J. Mdv1p is a WD repeat protein that interacts with the dynamin-related GTPase, Dnm1p, to trigger mitochondrial division. J. Cell Biol. 151, 353366 (2000).
5. 6.
7.
8.
9.
10.
11.
12.
13.
14.
15. Pilling, A. D., Horiuchi, D., Lively, C. M. & Saxton, W. M. Kinesin-1 and dynein are the primary motors for fast transport of mitochondria in Drosophila motor axons. Mol. Biol. Cell 17, 20572068 (2006). 16. Morris, R. L. & Hollenbeck, P. J. The regulation of bidirectional mitochondrial transport is coordinated with axonal outgrowth. J. Cell Sci. 104, 917927 (1993). 17. Chang, D. T., Honick, A. S. & Reynolds, I. J. Mitochondrial trafficking to synapses in cultured primary cortical neurons. J. Neurosci. 26, 70357045 (2006). 18. Chada, S. R. & Hollenbeck, P. J. Nerve growth factor signaling regulates motility and docking of axonal mitochondria. Curr. Biol. 14, 12721276 (2004). 19. Mannella, C. A. Structure and dynamics of the mitochondrial inner membrane cristae. Biochim. Biophys. Acta 1763, 542548 (2006). 20. Scorrano, L. et al. A distinct pathway remodels mitochondrial cristae and mobilizes cytochrome c during apoptosis. Dev. Cell 2, 5567 (2002). 21. Gilkerson, R. W., Selker, J. M. & Capaldi, R. A. The cristal membrane of mitochondria is the principal site of oxidative phosphorylation. FEBS Lett. 546, 355358 (2003). 22. Vogel, F., Bornhovd, C., Neupert, W. & Reichert, A. S. Dynamic subcompartmentalization of the mitochondrial inner membrane. J. Cell Biol. 175, 237247 (2006). 23. Wurm, C. A. & Jakobs, S. Differential protein distributions define two sub-compartments of the mitochondrial inner membrane in yeast. FEBS Lett. 580, 56285634 (2006). 24. Hermann, G. J. et al. Mitochondrial fusion in yeast requires the transmembrane GTPase Fzo1p. J. Cell Biol. 143, 359373 (1998). 25. Shaw, J. M. & Nunnari, J. Mitochondrial dynamics and division in budding yeast. Trends Cell Biol. 12, 178184 (2002). 26. Meeusen, S., McCaffery, J. M. & Nunnari, J. Mitochondrial fusion intermediates revealed in vitro. Science 305, 17471752 (2004). This study described an in vitro fusion assay that identifies mitochondrial fusion intermediates. 27. Detmer, S. A. & Chan, D. C. Complementation between mouse Mfn1 and Mfn2 protects mitochondrial fusion defects caused by CMT2A disease mutations. J. Cell Biol. 176, 405414 (2007). 28. Chen, H., Chomyn, A. & Chan, D. C. Disruption of fusion results in mitochondrial heterogeneity and dysfunction. J. Biol. Chem. 280, 2618526192 (2005). 29. Koshiba, T. et al. Structural basis of mitochondrial tethering by mitofusin complexes. Science 305, 858862 (2004).

REVIEWS
References 4244 showed the power of yeast genetic screens for identifying essential components of the mitochondrial fission machinery. Griffin, E. E., Graumann, J. & Chan, D. C. The WD40 protein Caf4p is a component of the mitochondrial fission machinery and recruits Dnm1p to mitochondria. J. Cell Biol. 170, 237248 (2005). Lee, Y. J., Jeong, S. Y., Karbowski, M., Smith, C. L. & Youle, R. J. Roles of the mammalian mitochondrial fission and fusion mediators Fis1, Drp1, and Opa1 in apoptosis. Mol. Biol. Cell 15, 50015011 (2004). Koch, A. et al. Dynamin-like protein 1 is involved in peroxisomal fission. J. Biol. Chem. 278, 85978605 (2003). Koch, A., Yoon, Y., Bonekamp, N. A., McNiven, M. A. & Schrader, M. A role for Fis1 in both mitochondrial and peroxisomal fission in mammalian cells. Mol. Biol. Cell 16, 50775086 (2005). Escobar-Henriques, M., Westermann, B. & Langer, T. Regulation of mitochondrial fusion by the F-box protein Mdm30 involves proteasome-independent turnover of Fzo1. J. Cell Biol. 173, 645650 (2006). Neutzner, A. & Youle, R. J. Instability of the mitofusin Fzo1 regulates mitochondrial morphology during the mating response of the yeast Saccharomyces cerevisiae. J. Biol. Chem. 280, 1859818603 (2005). Karbowski, M., Neutzner, A. & Youle, R. J. The mitochondrial E3 ubiquitin ligase MARCH5 is required for Drp1 dependent mitochondrial division. J. Cell Biol. 178, 7184 (2007). Nakamura, N., Kimura, Y., Tokuda, M., Honda, S. & Hirose, S. MARCH-V is a novel mitofusin 2- and Drp1-binding protein able to change mitochondrial morphology. EMBO Rep. 7, 10191022 (2006). Eura, Y., Ishihara, N., Oka, T. & Mihara, K. Identification of a novel protein that regulates mitochondrial fusion by modulating mitofusin (Mfn) protein function. J. Cell Sci. 119, 49134925 (2006). Wasiak, S., Zunino, R. & McBride, H. M. Bax/Bak promote sumoylation of DRP1 and its stable association with mitochondria during apoptotic cell death. J. Cell Biol. 177, 439450 (2007). Taguchi, N., Ishihara, N., Jofuku, A., Oka, T. & Mihara, K. Mitotic phosphorylation of dynamin-related GTPase Drp1 participates in mitochondrial fission. J. Biol. Chem. 282, 1152111529 (2007). Meisinger, C. et al. The morphology proteins Mdm12/ Mmm1 function in the major -barrel assembly pathway of mitochondria. EMBO J. 26, 22292239 (2007). Guo, X. et al. The GTPase dMiro is required for axonal transport of mitochondria to Drosophila synapses. Neuron 47, 379393 (2005). Stowers, R. S., Megeath, L. J., Gorska-Andrzejak, J., Meinertzhagen, I. A. & Schwarz, T. L. Axonal transport of mitochondria to synapses depends on milton, a novel Drosophila protein. Neuron 36, 10631077 (2002). Glater, E. E., Megeath, L. J., Stowers, R. S. & Schwarz, T. L. Axonal transport of mitochondria requires milton to recruit kinesin heavy chain and is light chain independent. J. Cell Biol. 173, 545557 (2006). Frederick, R. L., McCaffery, J. M., Cunningham, K. W., Okamoto, K. & Shaw, J. M. Yeast Miro GTPase, Gem1p, regulates mitochondrial morphology via a novel pathway. J. Cell Biol. 167, 8798 (2004). Sesaki, H., Southard, S. M., Yaffe, M. P. & Jensen, R. E. Mgm1p, a dynamin-related GTPase, is essential for fusion of the mitochondrial outer membrane. Mol. Biol. Cell 14, 23422356 (2003). Olichon, A. et al. Loss of OPA1 perturbates the mitochondrial inner membrane structure and integrity, leading to cytochrome c release and apoptosis. J. Biol. Chem. 278, 77437746 (2003). Amutha, B., Gordon, D. M., Gu, Y. & Pain, D. A novel role of Mgm1p, a dynamin-related GTPase, in ATP synthase assembly and cristae formation/ maintenance. Biochem. J. 381, 1923 (2004). Frezza, C. et al. OPA1 controls apoptotic cristae remodeling independently from mitochondrial fusion. Cell 126, 177189 (2006). Paumard, P. et al. The ATP synthase is involved in generating mitochondrial cristae morphology. EMBO J. 21, 221230 (2002). Minauro-Sanmiguel, F., Wilkens, S. & Garcia, J. J. Structure of dimeric mitochondrial ATP synthase: novel F0 bridging features and the structural basis of mitochondrial cristae biogenesis. Proc. Natl Acad. Sci. USA 102, 1235612358 (2005). 67. Dudkina, N. V., Heinemeyer, J., Keegstra, W., Boekema, E. J. & Braun, H. P. Structure of dimeric ATP synthase from mitochondria: an angular association of monomers induces the strong curvature of the inner membrane. FEBS Lett. 579, 57695772 (2005). 68. Messerschmitt, M. et al. The inner membrane protein Mdm33 controls mitochondrial morphology in yeast. J. Cell Biol. 160, 553564 (2003). 69. John, G. B. et al. The mitochondrial inner membrane protein mitofilin controls cristae morphology. Mol. Biol. Cell 16, 15431554 (2005). 70. Hobbs, A. E., Srinivasan, M., McCaffery, J. M. & Jensen, R. E. Mmm1p, a mitochondrial outer membrane protein, is connected to mitochondrial DNA (mtDNA) nucleoids and required for mtDNA stability. J. Cell Biol. 152, 401410 (2001). 71. Dimmer, K. S., Jakobs, S., Vogel, F., Altmann, K. & Westermann, B. Mdm31 and Mdm32 are inner membrane proteins required for maintenance of mitochondrial shape and stability of mitochondrial DNA nucleoids in yeast. J. Cell Biol. 168, 103115 (2005). 72. Chen, H., McCaffery, J. M. & Chan, D. C. Mitochondrial fusion protects against neurodegeneration in the cerebellum. Cell 130, 548562 (2007). This study showed that mitochondrial fusion is essential for respiratory function and that it protects neurons in the cerebellum from degeneration findings that will probably be relevant for understanding CMT2A pathogenesis. 73. Legros, F., Malka, F., Frachon, P., Lombes, A. & Rojo, M. Organization and dynamics of human mitochondrial DNA. J. Cell Sci. 117, 26532662 (2004). 74. Alavi, M. V. et al. A splice site mutation in the murine Opa1 gene features pathology of autosomal dominant optic atrophy. Brain 130, 10291042 (2007). 75. Davies, V. J. et al. Opa1 deficiency in a mouse model of autosomal dominant optic atrophy impairs mitochondrial morphology, optic nerve structure and visual function. Hum. Mol. Genet. 16, 13071318 (2007). 76. Labrousse, A. M., Zappaterra, M. D., Rube, D. A. & van der Bliek, A. M. C. elegans dynamin-related protein DRP-1 controls severing of the mitochondrial outer membrane. Mol. Cell 4, 815826 (1999). 77. Waterham, H. R. et al. A lethal defect of mitochondrial and peroxisomal fission. N. Engl. J. Med. 356, 17361741 (2007). This study provided clinical evidence for the essentiality of mitochondrial fission. 78. Verstreken, P. et al. Synaptic mitochondria are critical for mobilization of reserve pool vesicles at Drosophila neuromuscular junctions. Neuron 47, 365378 (2005). 79. Campello, S. et al. Orchestration of lymphocyte chemotaxis by mitochondrial dynamics. J. Exp. Med. 203, 28792886 (2006). 80. Arnoult, D. Mitochondrial fragmentation in apoptosis. Trends Cell Biol. 17, 612 (2007). 81. Youle, R. J. & Karbowski, M. Mitochondrial fission in apoptosis. Nature Rev. Mol. Cell Biol. 6, 657663 (2005). 82. Frank, S. et al. The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev. Cell 1, 515525 (2001). This study provided the first evidence that mitochondrial fission has a significant role in apoptosis. 83. Parone, P. A. et al. Inhibiting the mitochondrial fission machinery does not prevent Bax/Bak-dependent apoptosis. Mol. Cell. Biol. 26, 73977408 (2006). 84. Abdelwahid, E. et al. Mitochondrial disruption in Drosophila apoptosis. Dev. Cell 12, 793806 (2007). 85. Goyal, G., Fell, B., Sarin, A., Youle, R. J. & Sriram, V. Role of mitochondrial remodeling in programmed cell death in Drosophila melanogaster. Dev. Cell 12, 807816 (2007). 86. Jagasia, R., Grote, P., Westermann, B. & Conradt, B. DRP-1-mediated mitochondrial fragmentation during EGL-1-induced cell death in C. elegans. Nature 433, 754760 (2005). 87. Karbowski, M., Norris, K. L., Cleland, M. M., Jeong, S. Y. & Youle, R. J. Role of Bax and Bak in mitochondrial morphogenesis. Nature 443, 658662 (2006). 88. Brooks, C. et al. Bak regulates mitochondrial morphology and pathology during apoptosis by interacting with mitofusins. Proc. Natl Acad. Sci. USA 104, 1164911654 (2007). 89. Goldstein, J. C., Waterhouse, N. J., Juin, P., Evan, G. I. & Green, D. R. The coordinate release of cytochrome c during apoptosis is rapid, complete and kinetically invariant. Nature Cell Biol. 2, 156162 (2000). 90. Cipolat, S. et al. Mitochondrial rhomboid PARL regulates cytochrome c release during apoptosis via OPA1-dependent cristae remodeling. Cell 126, 163175 (2006). 91. Germain, M., Mathai, J. P., McBride, H. M. & Shore, G. C. Endoplasmic reticulum BIK initiates DRP1-regulated remodelling of mitochondrial cristae during apoptosis. EMBO J. 24, 15461556 (2005). 92. Alexander, C. et al. OPA1, encoding a dynamin-related GTPase, is mutated in autosomal dominant optic atrophy linked to chromosome 3q28. Nature Genet. 26, 211215 (2000). 93. Delettre, C. et al. Nuclear gene OPA1, encoding a mitochondrial dynamin-related protein, is mutated in dominant optic atrophy. Nature Genet. 26, 207210 (2000). References 92 and 93 showed that dominant optic atrophy is caused by mutations in OPA1. 94. Ferre, M., Amati-Bonneau, P., Tourmen, Y., Malthiery, Y. & Reynier, P. eOPA1: an online database for OPA1 mutations. Hum. Mutat. 25, 423428 (2005). 95. Olichon, A. et al. Effects of OPA1 mutations on mitochondrial morphology and apoptosis: relevance to ADOA pathogenesis. J. Cell Physiol. 211, 423430 (2006). 96. Kim, J. Y. et al. Mitochondrial DNA content is decreased in autosomal dominant optic atrophy. Neurology 64, 966972 (2005). 97. Lodi, R. et al. Deficit of in vivo mitochondrial ATP production in OPA1-related dominant optic atrophy. Ann. Neurol. 56, 719723 (2004). 98. Griparic, L., van der Wel, N. N., Orozco, I. J., Peters, P. J. & van der Bliek, A. M. Loss of the intermembrane space protein Mgm1/OPA1 induces swelling and localized constrictions along the lengths of mitochondria. J. Biol. Chem. 279, 1879218798 (2004). 99. Zuchner, S. & Vance, J. M. Mechanisms of disease: a molecular genetic update on hereditary axonal neuropathies. Nature Clin. Pract. Neurol. 2, 4553 (2006). 100. Zuchner, S. et al. Mutations in the mitochondrial GTPase mitofusin 2 cause Charcot-Marie-Tooth neuropathy type 2A. Nature Genet. 36, 449451 (2004). This study showed that mutations in MFN2 cause the peripheral neuropathy CMT2A. 101. Zuchner, S. et al. Axonal neuropathy with optic atrophy is caused by mutations in mitofusin 2. Ann. Neurol. 59, 276281 (2006). 102.Chung, K. W. et al. Early onset severe and late-onset mild Charcot-Marie-Tooth disease with mitofusin 2 (MFN2) mutations. Brain 129, 21032118 (2006). 103.Verhoeven, K. et al. MFN2 mutation distribution and genotype/phenotype correlation in Charcot-Marie-Tooth type 2. Brain 129, 20932102 (2006). 104.Baloh, R. H., Schmidt, R. E., Pestronk, A. & Milbrandt, J. Altered axonal mitochondrial transport in the pathogenesis of Charcot-Marie-Tooth disease from mitofusin 2 mutations. J. Neurosci. 27, 422430 (2007). 105.Niemann, A., Ruegg, M., La Padula, V., Schenone, A. & Suter, U. Ganglioside-induced differentiation associated protein 1 is a regulator of the mitochondrial network: new implications for Charcot-Marie-Tooth disease. J. Cell Biol. 170, 10671078 (2005).
45.
46.
47. 48.
49.
50.
51.
52.
53.
54.
55.
56.
57. 58.
59.
60.
61.
Acknowledgements
62.
This work was supported by grants from the National Institutes of Health. D.C.C. is an Ellison Medical Foundation Senior Scholar in Aging.
DATABASES
Entrez Gene: http://www.ncbi.nlm.nih.gov/entrez/query. fcgi?db=gene BAK | BAX | Dnm1 | DRP1 | FIS1 | FZO | Fzo1 | GDAP1 | Gem1 | MARCH5 |MDM10 | MDM12 | Mdm30 | Mdm31 | Mdm32 | Mdm33 | MFN1 | MFN2 | Mgm1 | milton | Miro | MMM1 | NGF | OPA1 | SMAC | TIM23 | Ugo1 OMIM: http://www.ncbi.nlm.nih.gov/entrez/query. fcgi?db=OMIM CMT2A | CMT4A
63.
64. 65. 66.
FURTHER INFORMATION
David C. Chans homepage: http://www.its.caltech.edu/~chanlab
all links are active in the online pDf

Enfermedades del mtDNA
ARTCULO
DE REVISIN
Enfermedades genticas del ADN mitocondrial humano

Abelardo Solano, Q. F. B.,(1) Ana Playn, Ph.D.,(1) Manuel J. Lpez-Prez, Ph.D.,(1) Julio Montoya, Ph.D.(1)
Solano A, Playn A, Lpez-Prez MJ, Montoya J. Enfermedades genticas del ADN mitocondrial humano. Salud Publica Mex 2001;43:151-161. El texto completo en ingls de este artculo est disponible en: http://www.insp.mx/salud/index.html Solano A, Playn A, Lpez-Prez MJ, Montoya J. Genetic diseases of the mitochondrial DNA. Salud Publica Mex 2001;43:151-161. The English version of this paper is available at: http://www.insp.mx/salud/index.html
Resumen Las enfermedades mitocondriales son un grupo de trastornos que estn producidos por un fallo en el sistema de fosforilacin oxidativa (sistema Oxphos), la ruta final del metabolismo energtico mitocondrial, con la consiguiente deficiencia en la biosntesis del trifosfato de adenosina (ATP, por sus siglas en ingls). Parte de los polipptidos que componen este sistema estn codificados en el cido desoxirribonucleico (DNA) mitocondrial y, en los ltimos aos, se han descrito mutaciones que se han asociado con sndromes clnicos bien definidos. Las caractersticas genticas del DNA mitocondrial, herencia materna, poliplasmia y segregacin mittica, confieren a estas enfermedades propiedades muy particulares. Las manifestaciones clnicas de estas enfermedades son muy heterogneas y afectan a distintos rganos y tejidos por lo que su correcto diagnstico implica la obtencin de datos clnicos, morfolgicos, bioqumicos y genticos. El texto completo en ingls de este artculo est disponible en: http://www.insp.mx/salud/ index.html Palabras clave: ADN mitocndrico; enfermedades mitocondriales; Espaa
Abstract Mitochondrial diseases are a group of disorders produced by defects in the oxidative phosphorylation system (Oxphos system), the final pathway of the mitochondrial energetic metabolism, resulting in a deficiency of the biosynthesis of ATP. Part of the polypeptide subunits involved in the Oxphos system are codified by the mitochondrial DNA. In the last years, mutations in this genetic system have been described and associated to well defined clinical syndromes. The clinical features of these disorders are very heterogeneous affecting, in most cases, to different organs and tissues and their correct diagnosis require precise clinical, morphological, biochemical and genetic data. The peculiar genetic characteristics of the mitochondrial DNA (maternal inheritance, polyplasmia and mitotic segregation) give to these disorders very distinctive properties. The English version of this paper is available at: http://www.insp.mx/salud/ index.html
Key words: DNA, mitochondrial; mitochondrial diseases; Spain
subcelulares L as mitocondriaselson organelosde las clulasque se encuentran en citoplasma eucariotas, cuya funcin principal es la produccin de la energa celular en forma de trifosfato de adenosina
(ATP, por sus siglas en ingls). Una de las particularidades de estos organelos es la de poseer un sistema gentico propio con toda la maquinaria necesaria para su expresin, es decir, para replicar, transcribir y
Este trabajo ha sido subvencionado por la Direccin General de Enseanza Superior e Investigacin Cientfica (PB97-1019), el Fondo de Investigaciones Sanitarias (FIS 98-0049-01), la Diputacin General de Aragn (P24/97) de Espaa, as como por el Consejo Nacional de Ciencia y Tecnologa (Conacyt) de Mxico. (1) Departamento de Bioqumica y Biologa Molecular y Celular, Universidad de Zaragoza, Zaragoza, Espaa. Fecha de recibido: 22 de mayo de 2000 Fecha de aprobado: 13 de noviembre de 2000 Solicitud de sobretiros: Dr. Julio Montoya. Departamento de Bioqumica y Biologa Molecular y Celular, Facultad de Veterinaria, Universidad de Zaragoza. Miguel Servet 177, E-50013 Zaragoza, Espaa. Correo electrnico: jmontoya@posta.unizar.es salud pblica de mxico / vol.43, no.2, marzo-abril de 2001 151
ARTCULO
DE REVISIN
Solano A y col.
traducir la informacin gentica que contiene. El cido desoxirribonucleico mitocondrial (mtDNA, por sus siglas en ingls) humano es una molcula circular compuesta por 16 569 pares de bases1 que contiene informacin para 37 genes: dos cidos ribonucleicos ribosmicos (rRNA), componentes de los ribosomas especficos mitocondriales, 22 de transferencia (tRNA), que son capaces de leer todo el cdigo gentico, y 13 polipptidos que forman parte de cuatro de los cinco complejos multienzimticos del sistema de fosforilacin oxidativa (sistema Oxphos), etapa terminal de la ruta de produccin de ATP. Estos pptidos corresponden a siete subunidades (ND1, 2, 3, 4, 4L, 5, 6) del dinucletido de nicotinamida y adenina reducido (NADH): ubiquinona xido-reductasa (complejo I); una subunidad (cyt b) de la ubiquinol: citocromo c xido-reductasa (complejo III); tres subunidades (CO I, II, III) de la citocromo c oxidasa (complejo IV), y dos subunidades de la ATP sintetasa (complejo V)2
OH Val rARN 16S rARN 12S Phe
BUCLE-D Thr Cyt. b
Cadena pesada
ND 1 lle F-met ND 2 Trp OL
>
CO
F IGURA 1. M APA
HUMANO . S E REPRESENTA LAS DOS HEBRAS DEL LOS GENES QUE CODIFICAN : RRNA
SEALADOS CON LA ABREVIATURA DEL AMINOCIDO QUE TRANSPORTAN, Y SECUENCIAS CODIFICADORAS DE PROTENAS
(CO:
SUBUNIDADES CITOCROMO C OXIDASA; CYT B:
ND: SUBUNIDADES DE NADH DESHIDROL INDICAN LOS LUGARES DE INICIACIN DE LA TRANSCRIPCIN DE LAS HEBRAS PESADA Y LIGERA, RESPECTIVAMENTE. OH Y O L SIMBOLIZAN LOS ORGENES DE
CITOCROMO B Y GENASA). H 1, H 2 Y REPLICACIN DE LA CADENA PESADA Y LIGERA
152
Leu
Pro
Glu ND 6 Gln Ala Asn Cys Tyr
ND 5
(figura 1). El resto de los polipptidos componentes de estos complejos, as como el complejo II completo, estn codificados en el DNA nuclear. La biognesis de este sistema constituye un caso nico en la clula ya que para su formacin se requiere la expresin coordinada de los dos sistemas genticos. Los caracteres moleculares bsicos y peculiares del sistema gentico mitocondrial se descubrieron al inicio de los aos ochenta,1,3-6 y en 1988 se encontraron las primeras mutaciones asociadas a enfermedades.7-9 Desde entonces, el nmero de mutaciones en el mtDNA y de enfermedades asociadas ha crecido de modo espectacular y ha generado lo que hoy se podra llamar como una medicina mitocondrial.10,11 Se designa con el nombre de enfermedades mitocondriales a un grupo de trastornos cuya caracterstica comn es un defecto en la produccin de ATP. Sin embargo, frecuentemente este trmino se aplica a trastornos producidos por daos en el sistema Oxphos, debido a que durante muchos aos slo se haban encontrado mutaciones en el mtDNA relacionados con los mismos. Hoy en da, se han comenzado a identificar genes nucleares codificantes de protenas de los complejos del sistema Oxphos o responsables de su ensamblaje. En este trabajo nos limitaremos a describir las enfermedades debidas a daos en el sistema gentico mitocondrial por ser las ms conocidas y por presentar un modo de herencia muy particular. Caracteres especficos de la gentica mitocondrial El tipo de herencia del sistema gentico mitocondrial, su localizacin en un organelo citoplasmtico, la disposicin contnua de los genes sin nucletidos intermedios ni intrones y la poliplasmia (alto nmero de copias en cada clula) proporcionan caracteres genticos que los diferencian claramente de los del DNA nuclear. Cada clula contiene entre unas 1 000 y 10 000 copias de mtDNA dependiendo del tejido, pasando por unos cuantos cientos en los espermatozoides y hasta unas 100 000 en el oocito. Cada mitocondria contiene entre 2 y 10 molculas. Herencia materna. El mtDNA se hereda por va materna con un patrn vertical no mendeliano. La madre trasmite su genoma mitocondrial a todos sus hijos, pero solamente las hijas lo pasarn a todos los miembros de la siguiente generacin y as sucesivamente. Esto se debe al elevado nmero de molculas de mtDNA que existe en los vulos (entre 100 000 y 200 000 copias) en comparacin con unos pocos cientos que hay en los espermatozoides. Adems, las mitocondrias
salud pblica de mxico / vol.43, no.2, marzo-abril de 2001
>
Cadena Ligera Ser
Asp
CO II
Lys
ATPasa 6 ATPasa 8
GENTICO DEL
DNA
DNA CON (12S Y 16S), TRNA,
Gly CO III

ND 4 ND 4L Arg ND 3
Leu Ser His
MITOCONDRIAL
ARTCULO
DE REVISIN
que puedan entrar en el vulo fecundado se eliminan por un proceso activo.12 Segregacin mittica. El fenotipo de una lnea celular puede variar durante la divisin celular debido a que las mitocondrias se distribuyen al azar entre las clulas hijas por lo que si en una clula coexisten dos poblaciones de mtDNA, una normal y otra mutada (heteroplasmia), a lo largo de las divisiones se podrn originar tres genotipos diferentes: homoplsmico para el DNA mitocondrial normal, homoplsmico para el DNA mutado y heteroplsmico. Por tanto, el fenotipo de una clula con heteroplasmia depender del porcentaje de DNA mutado que contenga. Si el nmero de molculas de mtDNA daado es relativamente bajo se produce una complementacin con las molculas de DNA normal y no se manifestar el defecto gentico. Cuando el DNA mutado sobrepasa un umbral determinado se manifestar un fenotipo patognico (efecto umbral), es decir, si la produccin de ATP llega a estar por debajo de los mnimos necesarios para el funcionamiento de los tejidos, debido a la produccin defectuosa de protenas codificadas en el mtDNA, se produce la aparicin de la enfermedad. El nmero de molculas de DNA es diferente en cada rgano y tejido segn la cantidad de energa requerida para su funcionamiento. Por ello, los tejidos que preferentemente se afectan son la visin, el sistema nervioso central, msculo esqueltico, corazn, islotes pancreticos, rin e hgado. Alta velocidad de mutacin. El mtDNA presenta una tasa de mutacin espontnea 10 veces superior a la del DNA nuclear. Este fenmeno puede estar causado porque en la mitocondria se producen continuamente radicales de oxgeno, como consecuencia de la oxidacin final de los compuestos carbonados, que pueden daar a un DNA que no est protegido por protenas. Debido a este hecho, la variacin de secuencias entre individuos de una misma especie es muy grande, hasta unos 70 nucletidos,13 y en un mismo individuo se estar generando, a lo largo de la vida, una pequea heterogeneidad en el mtDNA. De este modo, se ha llegado a proponer que la disminucin en la capacidad respiratoria de los tejidos que tiene lugar en el envejecimiento pueda ser debida a una acumulacin de este dao mitocondrial. 14 Esta teora tiene su primera evidencia en un trabajo del grupo de Attardi, que documenta que las mitocondrias se deterioran con la edad como consecuencia de la acumulacin de mutaciones.15 Las variaciones de secuencia existentes entre diferentes individuos han resultado muy tiles para estudios antropolgicos, etnolgicos y forenses, y es la base de la hiptesis de
que el hombre desciende de una mujer que vivi en Africa hace unos 250 000 aos (Eva mitocondrial).16 Enfermedades genticas del DNA mitocondrial Las enfermedades originadas por daos en el genoma mitocondrial tienen en comn el estar producidas por una deficiencia en la biosntesis de ATP, ya que toda la informacin que contiene este DNA est dirigida a la sntesis de protenas componentes del sistema Oxphos. Las manifestaciones de estas enfermedades son muy variadas y pueden afectar a todos los rganos y tejidos, ya que la sntesis de ATP se produce en todos ellos y a cualquier edad. Estas pueden presentar una serie de aspectos clnicos, morfolgicos y bioqumicos muy concretos que dan lugar a sndromes bien caracterizados pero, en la mayor parte de los casos, principalmente en edad peditrica, los sntomas son muy poco informativos y es slo la presencia de anormalidades neurolgicas, a veces acompaadas de aumento de cido lctico y de otros sntomas clnicos secundarios que afectan a diversos rganos, lo que da alguna orientacin en el diagnstico de una enfermedad mitocondrial.17 Entre las manifestaciones clnicas ms comunes se encuentran una o varias de las siguientes: desrdenes motores, accidentes cerebrovasculares, convulsiones, demencia, intolerancia al ejercicio, ptosis, oftalmoplejia, retinopata pigmentaria, atrofia ptica, ceguera, sordera, cardiomiopata, disfunciones hepticas y pancreticas, diabetes, defectos de crecimiento, anemia sideroblstica, pseudo obstruccin intestinal, nefropatas, acidosis metablica y otras ms secundarias. La presencia de uno o ms de estos sntomas requiere a continuacin de un estudio morfolgico, histoqumico y bioqumico para asegurar la naturaleza de estas enfermedades. As, con mucha frecuencia se encuentran: fibras rojo-rasgadas (acumulacin de mitocondrias anormales en tamao y nmero) en biopsias musculares teidas con tricromo de Gomori y fibras no reactivas a la tincin histoqumica de la citocromo c oxidasa; defectos en uno o varios complejos de la cadena respiratoria; y desarreglos metablicos con elevacin de lactato, piruvato o una aminoaciduria generalizada causados por una disfuncin de la cadena respiratoria que conlleva un aumento de equivalentes reductores en la mitocondria y citoplasma, y una alteracin del funcionamiento del ciclo de Krebs debido al exceso de NADH, lo que provoca una acumulacin de piruvato y su posterior conversin a lactato que difunde a la sangre. Sin em-
153
ARTCULO
DE REVISIN
Solano A y col.
bargo, la ausencia de algunos de estos caracteres no debe descartar la posibilidad de enfermedad mitocondrial, especialmente en pacientes en edad peditrica. Adems, los estudios familiares pueden ser decisivos si se comprueba la existencia de herencia materna de la enfermedad. El estudio gentico del paciente y familiares relacionados por va materna pueden asegurar finalmente que nos encontramos ante este tipo de trastornos. De hecho, hoy en da, el desarrollo y rapidez de las tcnicas de gentica molecular permiten, en ocasiones, una confirmacin de la enfemedad antes de haber realizado muchas de las pruebas anteriormente citadas. La complejidad del diagnstico de estas enfermedades hace preciso que los pacientes tengan que acudir a centros muy especializados donde se pueda llevar a cabo evaluaciones clnicas, metablicas, patolgicas, bioqumicas y genticas, y a que en su diagnstico estn implicados especialistas de muy diverso origen. Desde que en 1988 se describieran las primeras enfermedades causadas por daos en el mtDNA,7-9 se han encontrado ms de 150 mutaciones (ms 100 deleciones y unas 50 mutaciones puntuales) asociadas a enfermedades humanas. El inters por su estudio ha crecido enormemente debido al gran aumento de pacientes diagnosticados con estos trastornos y a que se presentan desde en recin nacidos hasta en adultos de todas las edades. Adems, muchas de estas mutaciones se trasmiten por lnea materna, como se ha indicado anteriormente, lo que hace que el diagnstico en un individuo pueda tener implicaciones en muchas generaciones de una familia. A pesar de la importancia que las enfermedades mitocondriales tienen ltimamente y de ser responsables de una considerable morbilidad, hasta ahora no se han realizado estudios exhaustivos sobre su prevalencia en la poblacin general. Las razones son mltiples:18 complejidad de las manifestaciones clnicas, necesidad de biopsias musculares para su diagnstico (no siempre se pueden detectar las mutaciones en muestras de sangre); necesidad de secuenciar todo el genoma mitocondrial para poder localizar mutaciones no detectadas hasta ahora, problemas ticos para realizar anlisis genticos presintomticos en nios, diagnstico errneo de muchos pacientes al no ser atendidos en centros especializados, etctera. Sin embargo, a pesar de todas estas dificultades, el grupo del doctor Turnbull, en Newcastle, Reino Unido, ha publicado muy recientemente los primeros datos epidemiolgicos de las enfermedades del mtDNA, centrados en la poblacin blanca de Europa del Norte residente en el noreste de Inglaterra.18 As, ha mostrado que los defectos en el mtDNA
154
son la causa de enfermedad en 6.57 de cada 100 000 individuos de la poblacin adulta trabajadora y que 7.59 por cada 100 000 adultos y nios no afectados corren el riesgo de desarrollar una de estas enfermedades. En total, 12.48 por 100 000 individuos (1 de cada 8 000) tienen o presentan un riesgo de padecer una enfermedad causada por daos en el mtDNA. Estos datos representan un mnimo de prevalencia porque, muy probablemente, el nmero de pacientes que han quedado sin diagnosticar es elevado por haber sido atendidos por mdicos de asistencia primaria, y no en clnicas neurolgicas, y que hayan podido pasar desapercibidos por presentar solamente algunos de los sntomas acompaantes de estas enfermedades como diabetes o ptosis. Los datos obtenidos por el grupo de Newcastle han permitido comprobar que la prevalencia de las enfermedades debidas a daos en el mtDNA, consideradas en su conjunto, es equivalente a la de otras enfermedades neurolgicas como la enfermedad de Huntington y la esclerosis amiotrfica lateral (6.4 y 6.2 por cada 100 000 individuos, respectivamente), y superior a la de otras enfermedades neuromusculares hereditarias como la distrofia de Duchenne (3.2 por cada 100 000 individuos).18 La experiencia de nuestro servicio de diagnstico en la Universidad de Zaragoza es que un 16% de los pacientes remitidos para el estudio gentico presentan una delecin o mutacin puntual.19,20,* No hemos realizado ningn estudio de lo que este nmero representa entre la poblacin general espaola; pero, seguro que tanto en el estudio realizado en Inglaterra como en nuestro laboratorio, el nmero de pacientes ser muy superior cuando se secuencie el mtDNA de todos los posibles implicados y se detecten nuevas mutaciones. Estos nmeros, junto al hecho de que no exista una terapia eficaz, y que, aunque algunas de estas enfermedades puedan mejorar o estabilizarse a lo largo de su curso, ilustran la importancia que tienen en relacin con la salud pblica, particularmente en cuanto a su atencin y consejo gentico, pues la mayora tiene un desenlace fatal. La heterogeneidad de las manifestaciones clnicas, morfolgicas y bioqumicas de las enfermedades del mtDNA, hace que su clasificacin se base muy frecuentemente en las caractersticas genticas de las mutaciones, a pesar de que, en algunos casos, una misma mutacin pueda dar lugar a fenotipos clnicos muy diversos. As, las enfermedades del mtDNA
* Solano A. Enfermedades del mtDNA (tesis doctoral). Zaragoza: Universidad de Zaragoza. En realizacin, 2000. salud pblica de mxico / vol.43, no.2, marzo-abril de 2001
ARTCULO
DE REVISIN
se pueden dividir en tres grandes grupos segn estn asociadas a mutaciones puntuales, a reorganizaciones o a disminucin de nmero de copias del mtDNA. En la severidad de la manifestacin de la enfermedad intervienen varios factores: la naturaleza de la mutacin, el grado de heteroplasmia, los requerimientos energticos del tejido y la capacidad del tejido para compensar el dao celular. A continuacin se presenta un resumen de las enfermedades ms comunes asociadas a estos tipos de mutaciones. Enfermedades asociadas a mutaciones puntuales en el mtDNA Dado el alto ndice de mutacin del mtDNA, como se ha indicado anteriormente, es posible encontrar un gran nmero de mutaciones puntuales. Sin embargo, la mayora son mutaciones silenciosas que no causan ningn tipo de defecto. Las mutaciones patolgicas se pueden encontrar tanto en los genes de tRNA, de rRNA, como en los codificantes de protenas, y responden siempre a un tipo de herencia materna. Neuropata ptica hereditaria de Leber. La neuropata ptica hereditaria de Leber (LHON) se caracteriza por la prdida bilateral de la visin central, originada por atrofia del nervio ptico. Aparece en la segunda o tercera dcada de la vida y afecta a ms hombres que a mujeres. Aunque normalmente slo la visin est afectada, hay casos en los que tambin aparecen trastornos en la conduccin cardiaca, neuropata perifrica y ataxia cerebelar. Esta fue la primera enfermedad humana de herencia materna que se asoci a una mutacin en el mtDNA. Despus se lleg a asociar hasta con 16 mutaciones puntuales (cuadro I), localizadas todas ellas en genes codificantes de protenas, y que se clasificaron en primarias, secundarias o intermedias segn su relacin con la aparicin de la enfermedad. Sin embargo, ltimamente slo tres, G3 460A, G11 778A y T14 484C, estn consideradas como primarias o patognicas verdaderas, siendo la G11 778A la responsable en 50% de los casos y la que provoca la forma ms severa de la enfermedad. Las tres se encuentran en genes que codifican algn polipptido del complejo I del sistema Oxphos. La deteccin de estas mutaciones se suele hacer en clulas sanguneas donde se encuentran tanto en forma homo como heteroplsmica. El resto de las mutaciones se consideran como secundarias, suelen acompaar a las anteriores en forma homoplsmica y se desconoce su relacin directa con la enfermedad. Entre estas ltimas vale la pena mencionar que la mutacin G15257A, consisalud pblica de mxico / vol.43, no.2, marzo-abril de 2001
derada como intermedia por algunos autores, se ha encontrado en varias familias analizadas en nuestro laboratorio por lo que pensamos que puede contribuir de forma decisiva a la aparicin de la enfermedad. La prevalencia de la enfermedad en hombres ha sugerido la influencia de un gen nuclear, y aunque se ha descrito un ligamiento de la enfermedad con el locus (DXS7) situado en el cromosoma X en familias finlandesas,21 no se ha podido confirmar en familias de otro origen. Sndrome de neuropata, ataxia y retinopata pigmentaria. Este sndrome est caracterizado por debilidad muscular neurognica, ataxia y retinitis pigmentosa. Suele ir acompaado de demencia, convulsiones y neuropata sensorial axonal, presenta una herencia materna y se ha asociado a una mutacin puntual, T8993G, en el gen de la subunidad 6 de la ATPasa (cuadro I). La mutacin aparece normalmente en forma heteroplsmica y en todos los tejidos estudiados: leucocitos, fibroblastos, msculo, rin y cerebro. Existe una alta correlacin entre la proporcin del DNA mutado y la severidad de la enfermedad. Sndrome de Leigh de herencia materna. El sndrome de Leigh de herencia materna (MILS) es una enfermedad muy heterognea que se puede presentar asociada a diferentes tipos de herencia, autosmica recesiva, ligada al cromosoma X o materna (mitocondrial) segn el gen que est daado. Es una enfermedad devastadora que se caracteriza por trastornos degenerativos multisistmicos que aparecen en el primer ao de vida, disfunciones del tallo cerebral y de los ganglios basales, desmielinizacin, regresin psicomotora, retraso en el desarrollo, ataxia, convulsiones, neuropata perifrica. El diagnstico se confirma por la presencia de lesiones necrticas cerebrales focales en el tlamo, tallo cerebral y ncleo dentado. La forma de la enfermedad, que se hereda por va materna, est producida por la mutacin en el gen de la subunidad 6 de la ATPasa, T8993G, la misma que produce el sndrome de neuropata, ataxia y retinopata pigmentaria, pero con un porcentaje de la mutacin superior a 90%. Otras formas menos severas de esta enfermedad se han asociado con un cambio TC* en la misma posicin del mtDNA. Sndrome de epilepsia mioclnica con fibras rojo-rasgadas (MERRF). Este sndrome de herencia materna, est caracterizado por epilepsia mioclnica, convulsiones generalizadas y miopata con presencia de fibras rojorasgadas. Otros sntomas clnicos que pueden acompaar a los anteriores son demencia, sordera, neuropata,
* Timina por citosina. 155
ARTCULO
DE REVISIN
Solano A y col.
atrofia ptica, fallo respiratorio y cardiomiopata. Aparece tanto en la infancia como en edad adulta y es de curso progresivo. Est asociado a la presencia de mutaciones en el gen del mtDNA para el tRNALys. En la mayora de los casos (80%-90%) se debe a una mutacin A8344G, pero tambin se han encontrado otras mi-
noritarias como T8356C (cuadro I), todas en forma heteroplsmica. El porcentaje de heteroplasmia necesario para la afectacin vara entre individuos jvenes (95%) e individuos por encima de los 60-70 aos (60%) del DNA mutado.13 La presencia de estas mutaciones en tRNA daa la sntesis de protenas.
Cuadro I
MUTACIONES EN EL MTDNA Y ENFERMEDADES ASOCIADAS

Enfermedad Mutacin en el mtDNA Gen afectado Referencias Enfermedad Mutacin en el mtDNA Gen afectado tRNALeu(uur) tRNALeu(uur) tRNAPro 12S rRNA tRNASer(ucn) tRNALeu(cun) tRNALys tRNALeu(uur) tRNAAsn tRNAAsn tRNALeu(uur) Citocromo b Citocromo b Citocromo b Citocromo b Citocromo b tRNAThr tRNALeu(uur) ATPase 6 ATPase 6 tRNALeu(uur) tRNALeu(uur) tRNALeu(uur) tRNATrp ND6 ND6 Referencias
53 54 55
LHON Mutaciones primarias
Mutaciones intermedias Mutaciones secundarias
NARP Leigh (MILS) MELAS
MERRF Diabetes y sordera Cardiomiopata (MICM)
G3460A G11778A T14484C G5244A G15257A T3394C T4160C T4216C A4917G G7444A T9101C G9438A G9804A G13708A G13730A G14459A G15812A T8993G T8993G T8993C A3243G C3256T T3271C T3291C T9957C A8344G T8356C A3243G A3260G C3303T A4269G A4300G A4317G C4320T G8363A T9997C
ND1 ND4 ND6 ND2 Citocromo b ND1 ND1 ND1 ND2 CO I ATPasa 6 CO III CO III ND5 ND5 ND6 cyt b ATPasa 6 ATPasa 6 ATPasa 6 tRNALeu(uur) tRNALeu(uur) tRNALeu(uur) tRNALeu(uur) COIII tRNALys tRNALys tRNALeu(uur) tRNALeu(uur) tRNALeu(uur) tRNAIle tRNAIle tRNAIle tRNAIle tRNALys tRNAGly
22,23 7 24 25 25 24 26 27 27 28 29 30 30 27 31 32 25 33 34,35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52
Miopata mitocondrial T3250C A3302G C15990T Sordera inducida por aminoglicsidos A1555G Sordera sensoneural T7445C Anemia sideroblstica G12301A Lipomatosis mltiple simtrica A8344G CPEO Delecin nica A3243G A5692G G5703A C3256T Intolerancia al ejercicio G15084A G15168A G15723A G14846A delecion de 24pb LIMM A15923G Muerte sbita A3251G Necrosis bilateral del estriado T9176C T8851C Multisistmicas A3251G A3252G C3256T Corea y demencia G5549A LHON y distona G14459A Diabetes y miopata T14709C Pearson Delecin nica Kearns-Sayre Delecin nica
56 57 58
59 60 61 62 63 63 64 64 64 64 64 65 66
67 68 66 69 63 70 32 71 72 73
LIMM: miopata mitocondrial infantil letal; LHON: neuropata ptica hereditaria de Leber; MELAS: encefalomiopata mitocondrial con acidosis lctica y episodios de accidentes cerebrovasculares; MERRF: epilepsia mioclnica con fibras rojo-rasgadas; MICM: Cardiomiopata de herencia materna; MILS: sndrome de Leigh de herencia materna; PEO: oftalmoplejia progresiva externa
156
ARTCULO
DE REVISIN
Sndrome de encefalomiopata mitocondrial con acidosis lctica y episodios de accidentes cerebro-vasculares (MELAS). Se trata de una encefalomiopata mitocondrial, de herencia materna, caracterizada por accidentes cerebrovasculares producidos a edad temprana que provocan una disfuncin cerebral subaguda y cambios en la estructura cerebral, y por acidosis lctica. Estos caracteres suelen ir acompaados de convulsiones generalizadas, dolor de cabeza, sordera, demencia y, a veces, presenta fibras rojo-rasgadas. Esta enfermedad ha sido asociada fundamentalmente con mutaciones en el gen del tRNALeu(UUR) del mtDNA. La mayor parte de los casos (80%) est asociada a la mutacin A3.243G, pero tambin se han encontrado otras con menor incidencia y alguna en genes codificantes de protenas (cuadro I), todas en forma heteroplsmica. Al igual que en la epilepsia mioclnica, las mutaciones en el tRNA daan la sntesis de protenas mitocondriales. Diabetes de herencia materna con sordera. Adems de los dos tipos clsicos de diabetes dependiente y no dependiente de insulina (tipo 1 y 2, respectivamente), se ha descrito recientemente un nuevo tipo de diabetes asociada a sordera, que no encuadra dentro de la clasificacin de la Organizacin Mundial de la Salud. Esta diabetes, de herencia materna, est producida por la mutacin A3.243G en el gen del tRNALeu(UUR) (cuadro I), la misma descrita para el sndrome de (MELAS). La frecuencia de diabetes y sordera es aproximadamente de un 1.5% de la poblacin diabtica total.74 Por otra parte, la diabetes es una de las enfermedades que se han descrito asociadas a otros sndromes mitocondriales como la encefalomiopata mitocondrial, oftalmoplejia progresiva externa crnica, KearnsSayre, Pearson y diabetes inspida, diabetes mellitus, atrofia ptica y sordera (DIDMOAD). Otras enfermedades del mtDNA asociadas a mutaciones puntuales Adems de las enfermedades descritas anteriormente, hay otras muchas que se han asociado a otras mutaciones puntuales (cuadro I). Entre ellas, se pueden citar las cardiomiopatas de herencia materna relacionadas fundamentalmente con mutaciones en el tRNAIle: la sordera inducida por aminoglicsidos que est producida por una mutacin en el rRNA 12S (A1555G), y otros tipos de sordera sindrmica o no sindrmica de herencia materna; LHON y distona; miopatas de herencia materna unidas a mutaciones en tRNA Leu, tRNAPro, tRNAAsn, tRNATyr; oftalmoplejia progresiva externa crnica; anemia sideroblstica; deficiencia fatal de la cadena respiratoria infantil; lipomatosis sisalud pblica de mxico / vol.43, no.2, marzo-abril de 2001
mtrica mltiple asociada a la mutacin A8.344G del gen del tRNALys (descrita en nuestro laboratorio) y, recientemente, se ha relacionado la intolerancia al ejercicio, como entidad propia, a mutaciones puntuales en el gen del citocromo b. As, se han descrito mutaciones en este gen que crean un codn de terminacin, que cambian un aminocido o, incluso, una delecin de 24 pares de bases. En el cuadro I se citan stos y otros sndromes que se han asociado a mutaciones puntuales y, sin ninguna duda, el espectro de fenotipos relacionados con mutaciones en el mtDNA aumentar ms en un futuro. Asimismo, cabe mencionar que alguna de las mutaciones, como la A3.243G, puede estar relacionada con muy diversos fenotipos clnicos como sindromes de encefalopata mitocondrial con acidosis lctica y episodios de accidentes cerebrovasculares de epilepsia mioclnica con fibras rojo-rasgadas y solapados, cardiomiopatas, CPEO, etctera. Actualmente, se estudia la posible implicacin del mtDNA en enfermedades neurodegenerativas como Parkinson y Alzheimer. Enfermedades asociadas a reorganizaciones en el DNA mitocondrial Adems de las mutaciones puntuales, el mtDNA puede sufrir otro tipo de daos como son la prdida de parte del mismo (deleciones) o la adicin de un nuevo fragmento del DNA (duplicaciones), que, como en los casos anteriores, afectan a la biognesis del sistema Oxphos y, por tanto, a la sntesis de ATP. En la actualidad hay descritos ms de 100 tipos de deleciones y slo unos cuantos casos de inserciones. Este tipo de mutaciones suelen ser espontneas, probablemente causadas por daos en genes nucleares que controlan la replicacin del mtDNA, aunque hay descritos casos de herencia materna.75 Se presentan siempre en forma heteroplsmica, ya que la homoplasmia sera incompatible con la vida, y se sabe que la gravedad de los casos aumenta con la edad debido a la ventaja replicativa de estas molculas de DNA ms pequeas en relacin con la de tamao normal. Los tres tipos de sndromes ms comunes en los que se presentan deleciones son los de Pearson, oftalmoplejia progresiva externa crnica y Kearns-Sayre. Sndrome de mdula sea-pncreas de Pearson. Es una enfermedad que aparece en los primeros aos de vida y que afecta a la hematopoyesis y a la funcin pancretica exocrina. Las caractersticas clnicas ms comunes son anemia sideroblstica con vacuolizacin de precursores de la mdula sea que se manifiesta con una anemia macroctica, trombocitopenia y neutropenia.
157
ARTCULO
DE REVISIN
Solano A y col.
Los nios afectados suelen morir antes de los tres aos de edad y los que sobreviven suelen desarrollar posteriormente el fenotipo de Kearns-Sayre, que veremos ms adelante. Estos pacientes presentan deleciones grandes nicas del mtDNA, en general son espordicas aunque se ha descrito algn caso de herencia materna. Oftalmoplejia progresiva externa crnica. Esta enfermedad est caracterizada por oftalmoplejia, ptosis bilateral de los prpados y miopata. Suele ir acompaada tambin de intolerancia al ejercicio y debilidad muscular. En el msculo se encuentran fibras rojo-rasgadas COX negativas. En general, es una enfermedad benigna que suele aparecer en la adolescencia o en adultos jvenes. Aparece de forma espordica sin historia familiar. Se ha asociado fundamentalmente a deleciones grandes y nicas en el mtDNA (ver ms adelante). Asimismo, se han encontrado otras formas de CPEO con mutaciones puntuales de herencia materna (cuadro I) o con deleciones mltiples de herencia autosmica recesiva o dominante. Sndrome de Kearns-Sayre. Este sndrome es, por otra parte, una enfermedad multisistmica progresiva caracterizada clinicamente por CPEO, retinopata pigmentaria atpica, ataxia, miopata mitocondrial, bloqueo de la conduccin cardiaca, elevados niveles de protena CSF (fluido cerebro espinal, por sus siglas en ingls), sordera y demencia. Aparece antes de los 20 aos de edad. Estas tres enfermedades estn causadas por deleciones (de 2 a 9 kb) en el mtDNA que suelen aparecer de forma espontnea. En general, la delecin es nica, pero tambin se han descrito casos de deleciones mltiples. La gravedad de la enfermedad depende del porcentaje de DNA mutado en el individuo. En general estn localizadas en el arco grande comprendido entre los orgenes de replicacin del DNA y mantienen siempre las secuencias requeridas para la replicacin del DNA y los promotores de la transcripcin. Entre todas las deleciones conocidas, hay una que aparece con ms frecuencia (hasta en 50%), la llamada delecin comn, que elimina un tramo de DNA de 4 977 pares de bases (entre los nucletidos 8 483 a 13 460), que comprende los genes localizados entre la subunidad 8 de la ATPasa y ND5 (figura 1). No existe una clara relacin entre el fenotipo y el tipo, tamao o porcentaje del DNA delecionado ya que la misma delecin puede dar lugar a varios fenotipos diferentes. La mayor parte de las deleciones encontradas estn flanqueadas por repeticiones directas de longitud variable (3-13 nt). Este hecho sugiere que la deleccin se produce por errores sucedidos en el proceso de replica158
cin dependientes de la presencia de estas repeticiones. La prdida de genes, especialmente la de los tRNA, hace que estos genomas no se puedan traducir y, por tanto, que sean dependientes de complementacin con molculas de mtDNA normales en la misma mitocondria. El umbral se suele alcanzar cuando el porcentaje de molculas delecionadas supera 60%. Existen otras enfermedades como una diabetes con sordera y atrofia ptica; miopatas en general; el sndrome de encefalomiopata mitocondrial neurogastrointestinal; el de diabetes mellitus, diabetes inspida, atrofia ptica y sordera, etctera, que estn asociadas a la presencia de deleciones en el mtDNA. Como se ha mencionado anteriormente, entre las reorganizaciones del mtDNA se pueden encontrar duplicaciones en pacientes con defectos en el sistema Oxphos. Estas pueden ser tambin espordicas o de herencia materna. Se han encontrado en pacientes de Kearns-Sayre, Pearson, diabetes mellitus, tubulopata renal y miopata mitocondrial e incluso en individuos normales. El mecanismo por el cual pueden causar la patogenicidad no est nada claro todava. Enfermedades asociadas a depleciones de DNA mitocondrial El tercer tipo de daos en el genoma mitocondrial que puede causar enfermedades no se debe a mutaciones propiamente dichas sino a una disminucin de los niveles del mtDNA. El espectro clnico que produce la deplecin es muy variado. Los casos descritos hasta ahora afectan fundamentalmente a nios con combinaciones variables de miopata, nefropata o hepatopata, miopata infantil fatal por fallo respiratorio y algn otro con implicacin multisistmica. La deplecin puede estar producida por mutaciones en genes nucleares que controlan el nmero de copias del mtDNA. Es, por tanto, un trastorno de herencia mendeliana que afecta a la coordinacin ncleo-mitocondria, y que parece ser autosmico recesivo. Regreso a la gentica mendeliana Debido al doble origen gentico nuclear y mitocondrial del sistema Oxphos las enfermedades genticas mitocondriales pueden estar originadas, adems de por mutaciones en genes del mtDNA con herencia materna, como ya hemos visto, por mutaciones en genes nucleares que codifican protenas mitocondriales, por mutaciones que afecten al procesamiento postraduccional, al importe de protenas por la mitocondria y al ensamblaje de los complejos, y por mutaciones que afecten al control nuclear del genoma
ARTCULO
DE REVISIN
mitocondrial, todas ellas con un tipo de herencia mendeliana. El primer caso ha sido hasta ahora el ms estudiado por estar el genoma mitocondrial completamente secuenciado y, por tanto, por la facilidad de encontrar mutaciones que afecten a los genes que codifica. Sin embargo, la mayor parte de los genes que componen el sistema Oxphos es de origen nuclear y cabe esperar que la mayora de las enfermedades causadas por la deficiencia de este sistema sean debidas a mutaciones en el DNA nuclear. As, por ejemplo, a pesar de que una de las causas del sndrome de Leigh sea una mutacin en el mtDNA de herencia materna, se sabe que se transmite con ms frecuencia por herencia autosmica recesiva, y se han encontrado e identificado mutaciones en genes de subunidades del complejo I, codificadas por el DNA nuclear76,77 y en el complejo II,78 codificado enteramente en el ncleo. Asimismo, se han localizado mutaciones en un gen nuclear (SURF 1) que codifica una protena que, aunque no forma parte del complejo IV, es necesaria para su ensamblaje.79,80 Adems, en la mitocondria existen muchas otras rutas metablicas en las que no participa para nada el mtDNA y cuya deficiencia puede causar encefalomiopatas mitocondriales. Por todo ello, la gentica mendeliana de las enfermedades mitocondriales est todava prcticamente por descubrir y nos proporcionar mucha informacin sobre estos trastornos.
Referencias
1. Anderson S, Bankier AT, Barrell BG, de-Bruijn MHL, Coulson AR, Drouin J et al. Sequence and organization of the human mitochondrial genome. Nature 1981;290:427-465. 2. Chomyn A, Mariottini P, Cleeter MWJ, Ragan CI, Doolittle RF, MatsunoYagi A et al. Functional assignment of the products of the unidentified reading frames of human mitochondrial DNA. En: Quagliarello E, Slater EC, Plamieri F, Saccone C, Kroon AM, ed. Amsterdam: Elsevier Sciences, 1985:259-275. 3. Montoya J, Ojala D, Attardi G. Distinctive features of the 5-terminal sequences of the human mitochondrial mRNAs. Nature 1981; 290: 465-470. 4. Ojala D, Montoya J, Attardi G. tRNA punctuation model of RNA processing in human mitochondria. Nature 1981; 290:470-474. 5. Montoya J, Christianson T, Levens D, Rabinowitz M, Attardi G. Identification of initiation sites for heavy strand and light strand trascription in human mitochondrial DNA. Proc Natl Acad Sci USA 1982; 79:7195-7199. 6. Montoya J, Gaines GL, Attardi G. The pattern of transcription of the human mitochondrial rRNA genes reveals two overlaping transcription units. Cell 1983; 34:151-159. 7. Wallace DC, Singh G, Lott MT, Hodge JA, Schurr TG, Lezza AMS et al. Mitochondrial DNA mutation associated with Lebers hereditary optic neuropathy. Science 1988; 242:1427-1430. salud pblica de mxico / vol.43, no.2, marzo-abril de 2001
8. Holt IJ, Harding AE, Morgan-Hughes JA. Deletions of muscle mitochondrial DNA in patients with mitochondrial myopathies. Nature 1988; 331:717-719. 9. Zeviani M, Moraes CT, DiMauro S, Nakase H, Bonilla E, Schon E et al. Deletions of mitochondrial DNA in Kearns-Sayre syndrome. Neurology 1988; 38:1339-1346. 10. DiMauro S, Bonilla E. Mitochondrial encephalomyopathies. En: Rosenberg RN, Prusiner SB, DiMauro S, Barchi RL, ed. Boston: ButterworthHeinemann, 1998:201-235. 11. Wallace DC. Mitochondrial DNA mutations and bioenergetic defects in aging and degenerative diseases. En: Rosenberg RN, Prusiner SB, DiMauro S, . Barchi RL, ed. Boston: Butterworth-Heinemann, 1998:237- 269. 12. Sutovsky P, Moreno RD Schatten G. Ubiquitin tag for sperm mitochondria. Nature 1999; 402:371. 13. Wallace DC. Diseases of the mitochondrial DNA. Annu Rev Biochem 1992; 61:1175-1212. 14. Miquel J. An update of the oxygen stress-mitochondrial mutation theory of aging: Genetic and evolutionary implications. Exp Gerontol 1998; 33:113-126. 15. Michikawa Y, Mazzucchelli F, Bresolin N, Scarlato G, Attardi G. Agingdependent large accumulation of point mutations in the human mtDNA control region for replication. Science 1999; 286:774-779. 16 Cann RL, Stoneking M, Wilson AC. Mitochondrial DNA and human evolution. Nature 1987; 325:31-36. 17. Munnich A, Rotig A, Chretien D, Cormier V, Bourgeron T, Bonnefont JP et al. Clinical presentation of mitochondrial disorders in childhood. J Inherited Metab Dis 1996;19:521-527. 18. Chinnery PF, Johnson MA, Wardell TM, SinghKler R, Hayes C, Brown DT et al. The epidemiology of pathogenic mitochondrial DNA mutations. Ann Neurol 2000; 48:188-193. 19. Beltrn B. Anlisis molecular de enfermedades del DNA mitocondrial: aplicacin al diagnstico (tesis doctoral). Zaragoza: Universidad de Zaragoza, 1995. 20. Playn A. Diagnstico molecular de enfermedades del DNA mitocondrial (tesis Doctoral). Zaragoza: Universidad de Zaragoza, 1999. 21. Vilkki J, Ott J, Savontaus ML, Aula P, Nikoskelainen EK. Optic atrophy in Leber hereditary optic neuroretinopathy is probably determined by an X-chromosomal gene closely linked to DXS7. Am J Hum Genet 1991; 48:486-491. 22. Howell N, Bindoff LA, Mccullough DA, Kubaccka I, Poulton J, Mackey D et al. Leber hereditary optic neuropathy: Identification of the same mitochondrial NDI mutation in six pedigrees. Am J Hum Genet 1991;49: 939-950. 23. Houponen K, Vilkki J, Aula P, Nikoskelainen EK, Savontaus ML. A new mitochondrial DNA mutation associated with Leber hereditary optic neuropathy. Am J Hum Genet 1991; 48:1147-1153. 24. Johns DR, Neufeld MJ, Park RD. An ND-6 mitochondrial DNA mutation associated with Leber hereditary optic neurtopathy. Biochem Biophys Res Commun 1992; 187:1551-1557. 25. Brown MD,Voljavec AS, Lott MT,Torrini A,Yang CC, Wallace DC. Mitochondrial DNA complex I and III mutations associated with Lebers hereditary optic neuropathy. Genetics 1992; 130:163-173. 26. Howell N, Kubacka I, Xu M, McCullogh DA. Leber hereditary optic neuropathy: Involvement of the mitochondrial NDI gene and evidence for an intragenic suppressor mutation. Am J Hum Genet 1991; 48:935-942. 27. Johns DR, Berman J. Alternative, simultaneous complex I mitochondrial DNA mutations in Lebers hereditary optic neuropathy. Biochem Biophys Res Commun 1991; 174:1324-1330. 28. Brown MD,Yang C, Trounce I, Torroni A, Lott MT, Wallace DC. A mitochondrial DNA variant, identified in Leber hereditary optic neuropathy patients, which extends the aminoacid sequence of cytochrome c oxidase subunit 1. Am J HumGenet 1992; 51:378-385. 29. Lamminen T, Majander A, Juvonen V, Wikstrom M, Aula P, Nikoskelainen E et al. A mitochondrial mutation at nt 9101 in the ATP synthase 6 gene 159
ARTCULO
DE REVISIN
Solano A y col.
associated with deficient oxidative phosphorylation in a family with Leber hereditary optic neuroretinopathy. Am J Hum Genet 1995; 56:1238-1240. 30. Johns DR, Neufeld MJ. Cytochrome-c oxidase mutations in Leber hereditary optic neuropathy. Biochem Biophys Res Commun 1993;196: 810-815. 31. Howell N, Halvorson S, Burns J, McCullough DA Poulton J. When does bilateral optical atrophy become Leber hereditary optic neuropathy? Am J Hum Genet 1994; 53:959-963. 32. Jun AS, Brown MD, Wallace DC. A mitochondrial DNA mutation at nucleotide pair 14459 of the NADH dehydrogenase subunit 6 gene associated with maternally inherited Leber hereditary optic neuropathy and dystonia. Proc Natl Acad Sci USA 1994; 91:6206-6210. 33. Holt IJ, Harding AE, Petty RKH, Morgan-Hughes JA. A new mitochondrial disease associated with mitochondrial DNA heteroplasmy. Am J Hum Genet 1990; 46:428-433. 34. Santorelli FM, Shanske S, Macaya A, Devivo DC, Dimauro S. The mutation at Nt 8993 of mitochondrial DNA is a common cause of Leighs syndrome. Ann Neurol 1993; 34:827-834. 35. Tatuch Y Robinson BH. The Mitochondrial DNA Mutation at 8993 Associated with NARP Slows the rate of ATP synthesis in isolated lymphoblast mitochondria. Biochem Biophys Res Commun 1993; 192:124-128. 36. Devries DD, Vanengelen BGM, Gabreels FJM, Ruitenbeek W, Vanoost BA. A 2nd missense mutation in the mitochondrial ATPase-6 gene in Leighs syndrome. Ann Neurol 1993; 34:410-412. 37. Goto YI, Nonaka I, Horai S. A mutation in the tRNALeu (UUR) gene associated with the MELAS subgroup of mitochondrial encephaloyopathies. Nature 1990; 348:651-653. 38. Sato W, Hayasaka K, Shoji Y, Takahashi T, Takada G, Saito M et al. A mitochondrial tRNA(Leu(uur) mutation at 3 256 associated with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Biochem Mol Biol Int 1994; 33:1055-1061. 39. Goto YI, Nonaka I, Horai S. A new mtDNA mutation associated with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Biochim Biophys Acta 1991; 1097:238-240. 40. Goto YI, Tsugane K, Tanabe Y, Nonaka I, Horai S. A new point mutation at nucleotide pair 3291 of the mitochondrial tRNA(Leu(uur)) gene in a patient with mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Biochem Biophys Res Commun 1994; 202:1624-1630. 41. Manfredi G, Schon EA, Moraes CT, Bonilla E, Berry GT, Sladky JT et al. A new mutation associated with MELAS is located in a mitochondrial DNA polypeptide-coding gene. Neuromuscular Disord 1995; 5:391-398. 42. Shoffner JM, Lott MT, Lezza AMS, Seibel P, Ballinger SW Wallace DC. Myoclonic epelipsy and ragged-red fiber desease (MERRF) is associated with a mitochondrial DNA tRNALYS mutation. Cell 1990; 61:931-937. 43. Silvestri G, Moraes CT, Shanske S, Oh SJ, DiMauro S. A new mtDNA mutation in the trans-RNA(Lys) gene associated with myoclonic epilepsy and ragged-red fibers (MERRF). Am J Hum Genet 1992; 51:1213-1217. 44. Van den Ouweland JMW, Lemkes HHPJ, Gerbitz KD, Maassen JA. Maternally inherited diabetes and deafness (MIDD): A distinct subtype of diabetes associated with mitochondrial tRNA (Leu(uur) gene point mutation. Muscle & Nerve 1995; S3:S124-S130. 45. Zeviani M, Gellera C, Antozzi C, Rimoldi M, Morandi L, Villani F et al. Maternal inherited myopathy and cardiomyopathy: Association with mutation in mitochondrial DNA tRNA (Leu(uur). Lancet 1991; 338:143-147. 46. Silvestri G, Santorelli FM, Shanske S,Whitley CB, Schimmenti LA, Smith SA et al. A new mtDNA mutation in the tRNA(Leu(uur)) gene associated with maternally inherited cardiomyopathy. Hum Mutat 1994; 3:37-43. 47. Taniike M, Fukushima H, Yanagihara I, Tsukamoto H, Tanaka J. Mitocondrial tRNAIle mutation in fatal cardiomypathy. Biochem Biophys Res Commun 1992; 186:47-53.
48. Casali C, Santorelli FM, Damati G, Bernucci P, Debiase L, DiMauro S. A novel mtDNA point mutation in maternally inherited cardiomyopathy. Biochem Biophys Res Commun 1995; 213:588-593. 49. Tanaka M, Ino H, Ohno K, Hattoti K, Sato W, Ozawa T et al. Mitochondrial mutation in fatal infantile cardiomyopathy. Lancet 1990;336:1452. 50. Santorelli FM, Mak SC,Vazquezacevedo M, Gonzalezastiazaran A, Ridaurasanz C, Gonzalezhalphen D et al. A novel mitochondrial DNA point mutation associated with mitochondrial encephalocardiomyopathy. Biochem Biophys Res Commun 1995; 216:835-840. 51. Santorelli FM, Mak SC, Elschahawi M, Casali C, Shanske S, Baram TZ et al. Maternally inherited cardiomyopathy and hearing loss associated with a novel mutation in the mitochondrial tRNA(Lys) gene (G8363A). Am J Hum Genet 1996; 58:933-939. 52. Merante F, Tein I, Benson L Robinson BH. Maternally inherited hypertrophic cardiomyopathy due to a novel T-to-C transition at nucleotide 9997 in the mitochondrial tRNA(glycine) gene. Am J Hum Genet 1994; 55:437-446. 53. Goto YI, Horai S, Matsuoka T, Yoga Y, Nihei K, Kobayashi M et al. Mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS): A correlative study of the clinical features and mitochondrial DNA mutation. Neurology 1992; 42:545-550. 54. Bindoff LA, Howell N, Poulton J, Mccullough DA, Morten KJ, Lightowlers RN et al. Abnormal RNA processing associated with a novel transfer RNA mutation in mitochondrial DNA - A potential disease mechanism. J Biol Chem 1993; 268:19559-19564. 55. Moraes CT, Ciacci F, Bonilla E, Ionasescu V, Schon EA Dimauro S. A mitochondrial transfer RNA anticodon swap associated with a muscle disease. Nat Genet 1993; 4:284-288. 56. Prezant TR, Agapian JV, Bohlman MC, Bu XD, Oztas S, Qiu WQ et al. Mitochondrial ribosomal RNA mutation associated with both antibioticinduced and non-syndromic deafness. Nat Genet 1993; 4:289-294. 57. Reid FM, Vernham GA, Jacobs HT. A novel mitochondrial point mutation in a maternal pedigree with sensorineural deafness. Hum Mutat 1994; 3:243-247. 58. Gattermann N, Retzlaff S, Wang YL, Berneburg M, Heinisch J, Wlaschek M et al. A heteroplasmic point mutation of mitochondrial tRNA(Leu)(CUN) in non-lymphoid haemopoietic cell lineages from a patient with acquired idiopathic sideroblastic anaemia. Br J Haematol 1996; 93:845-855. 59. Gamez J, Playn A, Andreu AL, Bruno C, Navarro C, Cervera C et al. Familial multiple symmetric lipomatosis associated with the A8344G mutation of mitochondrial DNA. Neurology 1998; 51:258-260. 60. Moraes CT, DiMauro S, Zeviani M, Lombes A, Shanske S Miranda AF. Mitochondrial DNA deletions in progressive external ophtalmoplegia and Kearns-Sayre syndrome. N Engl J Med 1989; 320:1293-1299. 61. Hammans SR, Sweeney MG, Hanna MG, Brockington M, Morganhughes JA, Harding AE. The mitochondrial DNA transfer RNA(Leu(uur)) A>G(3243) mutation-A clinical and genetic study. Brain 1995; 118:721-734. 62. Seibel P, Lauber J, Klopstock T, Marsac C, Kadenbach B, Reichmann H. Chronic progressive external ophthalmoplegia is associated with a novel mutation in the mitochondrial tRNA(Asn) gene. Biochem Biophys Res Commun 1994; 204:482-489. 63. Moraes CT, Ciacci F, Bonilla E, Jansen C, Hirano M, Rao N et al. Two novel pathogenic mitochondrial DNA mutations affecting organelle number and protein synthesis-Is the transfer RNA(Leu(uur) gene an etiologic hot spot? J Clin Invest 1993; 92:2906-2915. 64. Andreu AL, Hanna MG, Reichmann H, Bruno C, Penn AS, Tanji K et al. Exercise intolerance due to mutations in the cytochrome b gene of mitochondrial DNA. N Engl J Med 1999; 341:1037-1044. 65.Yoon KL, Aprile JR, Ernst SG. Mitochondrial tRNAThr mutation in fatal infantile respiratory enzyme deficiency. Biochem Biophys Res Commun 1991; 176:1112-1115.
160
ARTCULO
DE REVISIN
66. Sweeney MG, Bundey S, Brockington M, Poulton KR,Winer JB, Harding AE. Mitochondrial myopathy associated with sudden death in young adults and a novel mutation in the mitochondrial DNA leucine transfer RNA(uur) gene. Q J Med 1993; 86:709-713. 67. Thyagarajan D, Shanske S, Vazquezmemije M, Devivo D Dimauro S. A novel mitochondrial ATPase 6 point mutation in familial bilateral striatal necrosis. Ann Neurol 1995; 38:468-472. 68. Demeirleir L, Seneca S, Lissens W, Schoentjes E, Desprechins B. Bilateral striatal necrosis with a novel point mutation in the mitochondrial ATPase 6 gene. Pediatr Neurol 1995; 13:242-246. 69. Morten KJ, Cooper JM, Brown GK, Lake BD, Pike D Poulton J. A new point mutation associated with mitochondrial encephalomyopathy. Hum Mol Genet 1993; 2:2081-2087. 70.- Nelson I, Hanna MG, Alsanjari N, Scaravilli F, Morganhughes JA, Harding AE. A new mitochondrial DNA mutation associated with progressive dementia and chorea: A clinical, pathological, and molecular genetic study. Ann Neurol 1995; 37:400-403. 71. Rotig A, Colonna M, Bonnefont JP, Blanche S, Fischer A, Saudubray JM et al. Mitochondria DNA deletion in Pearsons marrow/pancreas sindrome. Lancet 1989; 902-903. 72. Hao HL, Bonilla E, Manfredi G, Dimauro S, Moraes CT. Segregation patterns of a novel mutation in the mitochondrial tRNA glutamic acid gene associated with myopathy and diabetes mellitus. Am J Hum Genet 1995; 56:1017-1025. 73. Zeviani M, Moraes CT, DiMauro S, Nakase H, Bonilla E, Schon EA et al. Deletions of mitochondrial DNA in Kearns-Sayre syndrome. Neurology 1998; 51:1525.
74. Maassen JA, Kadowaki T. Maternally inherited diabetes and deafness: A new diabetes subtype. Diabetologia 1996; 39:375-382. 75. Casademont J, Barrientos A, Cardellach F, Rotig A, Grau JM, Montoya J et al. Multiple deletions of mtDNA in two brothers with sideroblastic anemia and mitochondrial myopathy and in their asymptomatic mother. Hum Mol Genet 1994; 3:1945-1949. 76. Triepels RH, van den Heuvel LP, Loeffen JL, Buskens CA, Smeets RJ, Rubio-Gozalbo ME et al. Leigh syndrome associated with a mutation in the NDUFS7 (PSST) nuclear encoded subunit of complex I. Ann Neurol 1999; 45:787-790. 77. Loeffen J, Smeitink A, Triepels R, Smeets R, Schuelke M, Sengers R et al. The first nuclear-encoded complex I mutation in a patient with Leigh syndrome. Am J Hum Genet 1998; 63:1598-1608. 78. Bourgerom T, Rustin P, Chretien D, Birch-Machin M, Bourgeois M, Viegas-Pequignot E et al. Mutation of a nuclear succinate dehydrogenase gene results in mitochondrial respiratory chain deficiency. Nat Genet 1995; 11:144-149. 79. Tiranti V, Hoertnagel K, Carrozzo R, Galimberti C, Munaro M, Granatiero M et al. Mutations of SURF-1 in Leigh disease associated with cytochrome c oxidase deficiency. Am J Hum Genet 1998; 63:1609-1621. 80. Zhu ZQ, Yao JB, Johns T, Fu K, DeBie I, MacMillan C et al. SURF 1, encoding a factor involved in the biogenesis of cytochrome c oxidase, is mutated in Leigh syndrome. Nat Genet 1998; 20:337-343.
161

Sem10Biol Mitocondriopatias

Cargado por

Información del documento

Título original

Derechos de autor

Formatos disponibles

Compartir este documento

Compartir o incrustar documentos

Opciones para compartir

¿Le pareció útil este documento?

¿Este contenido es inapropiado?

Copyright:

Formatos disponibles

Sem10Biol Mitocondriopatias

Cargado por

Copyright:

Formatos disponibles

SEMINARIO SEMANA N 10 BIOLOGIA CELULAR Y MOLECULAR FACULTAD DE MEDICINA HUMANA

ADN MITOCONDRIAL. APLICACIONES EN MEDICINA FORENSE. MITOCONDRIOPATAS

Functions and dysfunctions of mitochondrial dynamics

870 | novEMBER 2007 | voluME 8

nATuRE REvIEWS | molecular cell biology

Low [ADP] High [ADP]

872 | novEMBER 2007 | voluME 8

Mitochondrial membrane potential

nATuRE REvIEWS | molecular cell biology

Ugo1 s-Mgm1 I-Mgm1 IMS OM

Mitochondrial F1F0 ATP synthase

874 | novEMBER 2007 | voluME 8

Adaptor poteins (Mdv1, Caf4)

nATuRE REvIEWS | molecular cell biology

Mitochondrial outer membrane permeabilization

876 | novEMBER 2007 | voluME 8

Table 1 | Disorders associated with mitochondrial perturbations

nATuRE REvIEWS | molecular cell biology

878 | novEMBER 2007 | voluME 8

64. 65. 66.

all links are active in the online pDf

nATuRE REvIEWS | molecular cell biology

voluME 8 | novEMBER 2007 | 879

Enfermedades del mtDNA

Enfermedades genticas del ADN mitocondrial humano

Key words: DNA, mitochondrial; mitochondrial diseases; Spain

OH Val rARN 16S rARN 12S Phe

BUCLE-D Thr Cyt. b

ND 1 lle F-met ND 2 Trp OL

SUBUNIDADES CITOCROMO C OXIDASA; CYT B:

Glu ND 6 Gln Ala Asn Cys Tyr

Cadena Ligera Ser

DNA CON (12S Y 16S), TRNA,

Leu Ser His

Enfermedades del mtDNA

salud pblica de mxico / vol.43, no.2, marzo-abril de 2001

Enfermedades del mtDNA

* Timina por citosina. 155

MUTACIONES EN EL MTDNA Y ENFERMEDADES ASOCIADAS

LHON Mutaciones primarias

Mutaciones intermedias Mutaciones secundarias

NARP Leigh (MILS) MELAS

MERRF Diabetes y sordera Cardiomiopata (MICM)

salud pblica de mxico / vol.43, no.2, marzo-abril de 2001

Enfermedades del mtDNA

Enfermedades del mtDNA

salud pblica de mxico / vol.43, no.2, marzo-abril de 2001

Enfermedades del mtDNA

salud pblica de mxico / vol.43, no.2, marzo-abril de 2001

También podría gustarte