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Regulacin de la expresin gentica

en eucariontes
Regulacin de la expresin gentica a
nivel TRANSCRIPCIONAL

Bibliografa: Alberts MBOTC Cap. 4 y 7


Activadores
Represores

Coactivadores
Factores de
transcripcin
basal
Promotor Basal: secuencia de nucletidos necesaria para la fijacin de la RNA
polimerasa.

Secuencias reguladoras:

A) Intensificadoras (enhancers): secuencias que estimulan la transcripcin y cuya


localizacin puede ser a miles de nucletidos de distancia "ro arriba o abajo" del
promotor

B) Silenciadoras (silencers): secuencias que inhiben la transcripcin. Tambin pueden


hallarse muy distantes del promotor.

Factores basales de transcripcin: complejo proteico que interacciona con el sitio


promotor. Son esenciales para la transcripcin pero no pueden aumentar o disminuir
su ritmo.
Factores especficos de la transcripcin: complejo de protenas reguladoras que
pueden ser activadoras o represoras.

A) Protenas activadoras: interaccionan con las secuencias intensificadoras del gen


(enhancers).

B) Protenas represoras: interaccionan con las secuencias silenciadoras del gen


(silencers).
Los factores de transcripcin son protenas de unin a DNA

Dominio de unin a DNA


Dominio transactivador
Domino regulable (p.ej. por hormona)
Interaccin de
los dominios de
unin a DNA con
sus secuencias
especficas

a) Hlice-vuelta-Hlice

b) Dedos de Zinc

c) Cierre de Leucina

d) Hlice-asa-Hlice
Receptores nucleares a hormonas (factores de
transcripcin especficos) y sus elementos de
respuesta en el DNA (Enhancer/Silencer)
Receptores a estrgenos, progesterona, testosterona
Receptores a glucocorticoides (cortisona, hidrocortisona, dexametasona)
Receptores a cido retinoico, tiroxina y Vitamina D
Relevancia del Enhancer y su Posicin respecto al Promotor Basal

Promotor Basal

Sin Promotor Basal

Con enhancer
Sin Promotor Basal

Con enhancer y
Promotor Basal

Con enhancer y
Promotor Basal

Con enhancer y
Promotor Basal

Con enhancer y
Promotor Basal
10.7 Chromatins role in eukaryotic gene regulation 323

(a) could become inactive. The three phenomena are X in-


Los diferentes estados
activation, imprinting, and position-effect variegation.
Short region of
DNA double helix
2 nm de la CROMATINA
MESSAGE The chromatin of eukaryotes is not uniform.
Highly condensed heterchromatic regions have fewer genes
Nucleosomes: and lower recombination frequencies than do the less
the basic unit 11 nm
condensed euchromatic regions.
of chromatin

X inactivation in female mammals


Chromatin fiber
of packed 30 nm In Chapter 15, you will learn about the effects of gene
nucleosomes copy number on the phenotype of an organism. For
now, it is sufficient to know that the number of tran-
(b)
scripts produced by a gene is usually proportional to the
number of copies of that gene in a cell. Mammals, for
example, are diploid and have two copies of each gene
located on their autosomes. However, as discussed in
Chapter 2, the number of the X and Y sex chromo-
somes differs between the sexes, with female mammals
having two X chromosomes to only one in males. The
mammalian X chromosome is thought to contain about
1000 genes. Females have twice as many copies of these
X-linked genes and would normally express twice as
much transcript from these genes as males. (Not having
a Y chromosome is not a problem for females, because
the very few genes on this chromosome are only re-
quired for the development of males.) This dosage im-
balance is corrected by a process called dosage compen-
sation, which makes the amount of most gene products
from the two copies of the X chromosome in females
Figure 10 - 29 The structure of chromatin. (a) The nucleosome equivalent to the single dose of the X chromosome in
in decondensed and condensed chromatin. (b) Chromatin males. This equivalency is accomplished by random inac-
La expresin gentica
en eucariontes requiere
de cambios en el estado
de la cromatina
Desacetilasas
de Histonas
(HDACs)

Complejos
remodeladores de
cromatina

Acetilasas de
Histonas
(HATs)
El cdigo de histonas (Modificaciones postraduccionales)
THEREGULATION
OFCHROMATIN
STRUCTURE 223

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r . q n a x a l s t R s s R A GL Q F P v G R V - i r H2A
1 5 9 1315

119

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A .A le
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l-r
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. "ffi'
5 12 141s 20 2324
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120
A.&
M$A':{.M
YY l? Y Ml M MIP Y Y
A * r K e r A R K S r G G K An * l o r x n o * l $ A p A r c GV K - K
2 4 9 r0 14 1718 " o 23 262728 36 79

AAA
PMA A M A M M
ttltttll
S,:Rl'XCCXI: L GKaIG1rKRHRKVLIT.DNT !G i L'-K
135812162079

N-terminaltails Sl"brbr.
domains

M methylation P phosphorylation acetvlation U ubiquitylation


bottom view I ffi I

(A) (B)

Figure4-39 The covalentmodificationof core histonetails.(A)Thestructureof the nucleosome highlightingthe location


(B\Well-documented
of the first30 aminoacidsin eachof its eight N-terminalhistonetails(green). modificationsof the
226
Cmo se interpreta
Chapter4: DNA,Chromosomes,
el cdigo de Histonas?
and Genomes

(A)
Figure 4-44 Somespecificmeanin
the histonecode.(A)The modifica
A
on the histoneH3 N-terminal tail a
M AAA M
MM MvrlM shown,repeatedfrom Figure4-39
lP I rl YI? M M
r (B)The H3 tail can be markedby di
RK KS K RK K RKS K K combinations of modifications tha
24 910 14 1118 2 3 262728 36 79 conveya specificmeaningto the s
of chromatinwherethis combinat
occurs.Onlya few of the meaning
known,includingthe four exampl
(B) modification state "meaning"
shown.Tofocuson just one examp
trimethylation of lysine9 attractsth
heterochromatin-specific proteinH
M whichinducesa spreadingwaveof
h e t e r o c h r o m a t i fno r m a t i o n , furtherlysine9 trimethylation follo
N
g e n es i l e n c i n g by furtherHP1binding,accordingt
9 generalschemethat will be illustr
MA shortly(seeFigure4-46). Not show
rl
g e n ee x p r e S s r o n the fact that,asjust implied(seeFi
KK 4-43),readingthe histonecodege
49 involvesthe joint recognitionof ma
PA othersiteson the nucleosome alon
t l
g e n ee x p r e S S t o n the indicatedH3 tail recognition. In
SK addition,specificlevelsof methyla
10 14 (mono-,di-,or tri-methylgroups)a
required,as in Figure4-42.
M
I
s i l e n c i n go f H o x g e n e s ,
K X c h r o m o s o m ei n a c t i v a t i o n
27

The marks on nucleosomes due to covalent additions to histones are


dynamic, being constantly removed and added at rates that depend on their
chromosomal locations. Because the histone tails extend outward from the
bonds.Thisis one of manyPHDdomains
protein complexes that execute an appropriate biological function at the right that recognizemethylatedlysineson
time (Figure 443). histones;differentdomainsbind tightly
to lysineslocatedat differentpositions,

Lectura del cdigo de


orotein modules scaffold betweena
and they can discriminate
b i n d i n gt o s p e c i f i c protein
h i s t o n em o d i f i c a t i o n s mono-,di-,and tri-methylated lysine.In a
similarway,othersmallproteinmodules
histonas
on nucteosome
recognize specifichistonesidechains
that havebeen markedwith acetyl
groups,phosphategrouPs,and so on.
(Adaptedfrom P.V.Penaet al.,Nature
03,2006.With permissionfrom
Transmisin de la
442:100-1
MacmillanPublishers Ltd.)

c o v al e n t
modification
informacin
o n h i s t o n et a i l
(mark)

C O D ER E A D E R
BINDS AND
ATTRACTS OTHER
COMPONENTS
p r o t e i nc o m p l e xw i t h
catalyticactivitiesand
a d d i t i o n a lb i n d i n gs i t e s

Figure4-43 Schematicdiagramshowing
how the histone code could be read by a
Remodelacin de
code-readercomplex.A largeprotein
complexthat containsa seriesof protein

cromatina
modules,eachof which recognizes a
specifichistonemark,is schematically
illustrated(green).This
"code-reader
complex"will bind tightlyonly to a region
of chromatinthat containsseveralof the

Activacin/
differenthistonemarksthat it recognizes'
Therefore,only a specificcombinationof
markswill causethe complexto bind to
Silenciamiento
chromatinand attractadditionalprotein
complexes(purple)thatcatalyzea

Transcripcional
biolooicalfunction.
somewhatconfusingbecausethey
Typically, a few relatively short stretches of nucleotide sequence guide the encompass an enormousvarietyof
assembly of a group of regulatory proteins on DNA (seeFigure 7-51). However, proteinsincludinghistonereadersand
in some extreme cases of regulation by committee a more elaborate writers,chromatinremodelingcomplexes,
and manyotherclasses of proteins.
Some
protein-DNA structure is formed (Figure 7-52). Since the final assembly but
haveno intrinsicactivitythemselves
requires the presence of many gene regulatory proteins that bind DNA, it pro- simplyserveasa "scaffolding"to attract
vides a simple way to ensure that a gene is expressedonlywhen the cell contains thosethat do.
the correct combination of these proteins. We saw earlier how the formation of
heterodimers in solution provides a mechanism for the combinatorial control of
gene expression. The assembly of complexes of gene regulatory proteins on
DNA provides a second important mechanism for combinatorial control, one
that offers far richer opportunities.

Compfex GeneticSwitchesThatRegulateDrosophilo
DevelopmentAre BuiltUp from SmallerModules
Given that gene regulatory proteins can be positioned at multiple sites along
long stretches of DNA, that these proteins can assembleinto complexes at each
site, and that the complexes influence the chromatin structure as well as the
recruitment and assembly of the general transcription machinery at the pro-
moter, there would seem to be almost limitless possibilities for the elaboration
of control devices to regulate eucaryotic gene transcription.
A particularly striking example of a complex, multicomponent genetic
switch is that controlling the transcription of the Drosophila Euen-skipped(Eue)
gene, whose expression plays an important part in the development of the
Drosophila embryo. If this gene is inactivated by mutation, many parts of the
embryo fail to form, and the embryo dies early in development. As discussedin
activates
Chapter 22, at the stage of development when Eue begins to be expressed,the +.
transcfl pI|on
embryo is a single giant cell containing multiple nuclei in a common cytoplasm.
This cytoplasm is not uniform, however: it contains a mixture of gene regulatory
proteins that are distributed unevenly along the length of the embryo, thus pro-
viding positional information that distinguishes one part of the embryo from
another (Figure 7-53). (The way these differencesare initially set up is discussed Figure7-52 Schematicdepictionof a
in Chapter 22.) Although the nuclei are initially identical, they rapidly begin to committee of gene regulatory proteins
bound to an enhancer.The protein
express different genes because they are exposed to different gene regulatory
shownin yellowiscalledan architectural
proteins. The nuclei near the anterior end of the developing embryo, for exam-
Secuencia de eventos
proteinsinceits mainroleis to bendthe
ple, are exposed to a set of gene regulatory proteins that is distinct from the set DNAto allowthe cooperative assembly
that influences nuclei at the posterior end of the embryo. of the othercomponents. Thestructure

en la activacin
The regulatory DNA sequencescontrolling the Eue geneare designedto read
the concentrations of gene regulatory proteins at each position along the length
deoictedhereis basedon that found in
the controlregionof the genethat codes
for a subunitof the T cellrecePtor

transcripcional
of the embryo and to interpret this information in such a way that the Eue gene (discussed in Chapter25),and it activates
is expressedin seven stripes, each initially five to six nuclei wide and positioned transcriptionat a nearbypromoter.Only
precisely along the anterior-posterior axis of the embryo (Figure 7-54). How is certaincellsof the developingimmune
this remarkable feat of information processing carried out? Although not all of system,which eventuallygive riseto
the molecular details are understood, several general principles have emerged matureT cells,havethe completeset of
from studies of Eue and other Drosophila genesthat are similarly regulated. Droteinsneededto form this structure.
Sealizacin para la activacin de los factores
de transcripcin especficos (CASO I)

El ligando que estimula


(cortisol) es liposoluble y
entra a la clula.

El GR se encuentra en
complejo inactivo con Hsp
y es liberado por el
ligando.

Entra a ncleo donde se


dimeriza y acta sobre sus
elementos de respuesta
estimulando la
transcripcin de genes
especficos.

Receptor a Glucocorticoides (GR)


Sealizacin para la activacin de los factores de
transcripcin especficos (CASO II)
Stat1

En este caso el
ligando NO entra a la
clula y su receptor es
una protena de
membrana.

Al unir el ligando, el
receptor se activa por
fosforilacin y acta
como cinasa
fosforilando a Stat1
en el citosol.

Esta fosforilacin
promueve la
dimerizacin y
entrada al ncleo del
factor transcripcional.
Los activadores pueden actuar de manera conjunta
Formas de accin de la protena activadora

Competencia con el Represor por el mismo sitio de unin


Formas de accin de la protena activadora

Enmascaramiento del dominio de transactivacin


Formas de accin de la protena activadora

Interaccin directa con factores de transcripcin


Complejos formados in situ sobre el DNA

Cada gen tiene una combinacin particular de intensificadores y


silenciadores.

Genes distintos pueden compartir idnticas secuencias


intensificadoras y silenciadoras, pero no existen dos genes que
posean la misma combinacin de estas secuencias reguladoras.
Regulacin en el desarrollo por protenas hometicas
(Embriones de Drosophila)

Activadores transcripcionales Represores transcripcionales


Expresin del gen Eve
La zona de regulacin involucra activadores y
represores

activadores

represores
En la zona de la banda 2 se dan las condiciones de
represor y activador
Regulacin de la expresin gentica a
nivel POST-TRANSCRIPCIONAL
Cambios en la expresin gentica a nivel TRADUCCIN

A B B B B
B B B B
Regulacin traduccin ! B B B B
B B B B
Durante el desarrollo embrionario de Drosophila
ocurre la traduccin selectiva de mRNAs en el embrin
para determinar la polaridad del cuerpo de la mosca

Traduccin Traduccin
activa inactiva

Traduccin
Traduccin inactiva
inactiva
En ausencia de HEMO
se activa una protena
cinasa que fosforila a
eIF2.

eIF2 fosforilado no es
capaz de intercambiar
el GDP por GTP, ya
que queda formando
complejo inactivo con el
eIF2B.

Hay deplecin de
complejo ternario e
inhibicin de la
traduccin.

Regulacin por grupo HEMO


en eritrocitos
Regulacin por siRNAs
Presencia de RNA de doble
cadena (por virus, RNA anti-
sentido, etc.)

Se generan siRNAs de 21 nt
por corte con DICER

Se forma RISC cargado con


siRNA y varias protenas
incluyendo ARGONAUTA

RISC dirige el siRNA a su


mRNA blanco con el cual
forma complementariedad
PERFECTA y promueve el
corte y degradacin de este
mRNA
Regulacin por miRNAs
Los miRNAs se transcriben por
la RNA polimerasa II como
transcritos primarios y son
procesados por DROSHA en el
ncleo

Luego son exportados al


citoplasma y son cortados por
DICER a tamao de 21 nt

Los miRNAs son tomados por


RISC con ARGONAUTA y son
dirigidos a sus mRNA blanco
con los cuales muestran
complementariedad
IMPERFECTA generalmente
en sus regiones 3UTR

RISC cargado con miRNAs


dirige la inhibicin traduccional
de los mRNAs blanco
RESUMEN

Control transcripcional A- Factores de transcripcin


B- Grado de condensacin de la cromatina
C- Grado de metilacin del ADN

Control procesamiento del Empalme alternativo


ARNm Grado de poliadenilacin

Control transporte del Mecanismos que determinan si el ARNm


ARNm maduro sale o no a citosol

Control traduccional Mecanismos que determinan si el ARNm


presente en el citosol es o no traducido

Control de la degradacin Mecanismos que determinan la supervivencia


del ARNm del ARNm en el citosol

Control de la actividad Mecanismos que determinan la activacin o


proteica inactivacin de una protena, as como el
tiempo de supervivencia de la misma.