Signal sequences govern genome rearragements

Several long segments of chromosomal DNA carrying clusters of v gene

segment, D gene segments, and j gene segments of both mice and humans
have now been sequenced, and the resulting nucleotide pair sequences
suggest the presence of specific v-j, v-d and d- j joining signals. The same signal
sequences are found adjacent to all v gene segment. Similarly. All j gene
segments have identical signal sequences located adjacent to their coding
sequences; however, their signal sequence is different from that adjacent to v
gene segments. Likewise, d and c gene segments have their own adjacent signal
sequences.
The signal sequences controlling V-J, V-D and D-J joining contain 7-base pair
(heptamer) and 9- base pair (nonamer)- long sequences separated by spacers of
different, but specific lengths. For Vk-Jk joining, the spacer in the Vk signal
sequence is 12 nucleotide pairs long, whereas that in the Jk signal sequence is
22 nucleotide pairs long. The heptamer and nonamer sequences located after
the Vk gene segments are complementary (with the exception of one base pair)
to those preceding the Jk gene segments.these signal sequences have the
potential to form stem and loop structures as diagrammed in fig 16.8 , thus
bringing the Vk and Jk gene segments into juxtaposition for joining. Apparently,
joining will occur only when one signal sequence contains a 12 base pair
spacer and the other contains a 22 base pair spacer. This requirement would
supposedly be enforced by the specific protein mediating the joining process.
Very similar signal sequences appear to control Vh D and D- Jh joining, whereas
somewhat different signal sequences mediate class switching
Antibody diversity variable joining sites and somatic mutations
Subsequent studies showed the much of this additional diversity could be
explained by variation in the exact site of recombination during the V- J joining
events.

How many combinations?

One can readily see that a large amount of diversity can be generated by the
joining of antibody gene segments as just described . for example, consider the
number of different kappa light chains possible in humans: 300 v gene
segmentsx 5 j segments = 1500 fused v j gene segments. The heavy chains
variable region provides even greater diversity because of the multiple D gene
segments. If there are 300 vhgene segments, 25 d gene segments and 6jh gene
segments in human germ line cells, 45000 different heavy chain variable regions
could be assembled. Using these estimates, 67,500,000 different antigen binding
sites could be produced using just kappa light chains. Lambda light chains
produce another level of diversity.
Clearly, these antibody gene segment fusions provide for a vast amount of
antibody diversity. We now know , however that further diversity is generated in
two additional ways. (1) somatic mutation and (2) variability in the sites at which
v-j, v-d, and d-j joining events occur. In total,the possible range of antibody
diversity seems almost limitless.

Regulations a tissue specific enhancer

It has been known for several years that germ line antibody gene are not
transcribed or are transcribed at very low levels. Yet, in antibody producing B
lymphocytes, 10 to 20 percent of the mRNA molecules are antibody gene
transcripts. What, then, is responsible for the activation of transcription of
antibody genes that undergo rearragements and become active? In the case of
the heavy chains gene , the answer appears to be that the rearragement process
brings the promoters located upstream of L11 V11 gene segments into the
range of influence of a strong enhancer element located in the intron between
the jh gene segment and the Chm gene segment (fig 16.10). each LH VH gene
segment contains an upstream promoter. However, prior to the genomic
rearragement events that lead to heavy chain synthesis, this enhancer is over
100,000 nucleotide pairs away from the closer LH-VH promoter. Presumably, this
enhancer cannot activate transcription from promoters that are located that far
away. However, rearragement events occuring during B cell differentiation (see
fig 16.5) move the promoter of the closest LH- VH gene segment to within less
than 2000 nucleotide pairs from the enhancer (fig 16.10, bottom). The enhancer
can now activate transcription from the promoter located upstream from the LH
VH gene segment. The enhancer involved in the activation of heavy chain
synthesis is tissue specific, it activates transcription only in lymphocytes and has
no effect in cells derived from other tissue. Presumably, the activation process
requires the presence of a transcriptional activating factor thatiss synthesized in
lymphocytes, but not in other types of cells.

A similar enhancer element has been found in the intron between the light chain
J gene segment cluster and the C coding sequence. Thus, it seems likely that the
movement of antibody gene promoters into the range of influence of tissue
spesific enhancers may be a general mechanism of antibody genes during the
differentiation of B lymphocytes.

Clonal selection
Kita tidak bisa menghindar tentang pertanyaan yaitu tentang bagaimana
suatu organisme menginisiasi sintesis dari antibodi yang spesific pada
antigentidak bisa sebelum melaluinya . berikut merupakan penjelasan dari clonal
selection theory mengingat pada semua antibodi memproduksi satu lymphocyte
B yang memiliki antigen sama yang terikat secara spesific. Tetapi ada perbedaan
sel yang berada di populasi lymphocyte B yang dapat mengalami perbedaan
urutan genome dari produksi antibodi yang berbeda spesificnya. Populasi dari
limfosit B pada manusia dan tikus akan memproduksi antibodi yang sangat
banyak macamnya.
Allelic exclusion
Consider one final point about the genetic control of antibody synthesis.
Each B lymphocyte makes only one type of antibody. Why?
Mammalian cells are diploid they carry two sets of genetic information coding for
each of the antibody chairs. But only one productive genome rearragement of

light chain coding sequences and one productive genome rearragement of heavy
chain coding sequences occur in each B lymphocyte! This phenomenon is called
allelic exclusion because one of the alleles is excluded from being expressed.
How? Why? At present. We still dont know. Clearly , there must be some type pf
a feedback mechanism that arrests the recombination process involved in these
antibody gene rearragements once a productive rearragements has occurred and
the cell has started to synthesize a functional antibody. The simplest mechanism
would involve inhibition of this process by the mature antibody it self. However ,
further work will be required to establish the mechanism by which allelic
exclusion occurs.
T cell receptor variability
T lymphocytes mediate the cellular immune response. The T cells recognize
antigens on the surface of cells and kill the cells carrying these antigens. Like the
antibodies produced by B lymphocytes, T cells can recognize and destroy cells
carrying anamazing variety of antigens. Thus, the T cell response also exhibits a
phenomenal degree of specificity. How is this specificity produced? The answer is
that T cells produce membrane bound receptors that are very similar to the
antibodies produced by B lymphocytes. Moreover, the diversity of T cell receptor
specificity is produced by genome rearragements analogous to those involved in
antibody production. But how do T lymphocytes avoid interacting with free
antigens to avoid duplicating the function of B cells in the immune response? As
it turns out, T cells must simultaneously recognize both the offending antigen on
the cell surface and another protein that occurs only attached to cell surfaces.
This second cell surface protein that the T cell must recognize is the product of
one of many genes in the major hostocompatibility complex (MHC). The MHC
locus encodes a complex group of proteins that are present on all the cells in the
body of a human (or a mouse). Thus , T cells are able to recognize and destroy
any cell that is producing a given antigen (e.g the coat protein of a virus) in any
tissues of the body. The interaction of the T cell receptor with the two types of
cell surface antigens (the offending antigen and the histocompatibility antigen)is
illustrated in fig 16.1.
The T cell receptors are composed of two polypeptide chains, a and B, each
encoded by L-V, D, J and C gene segment, is just like antibody chains, contain
variable regions that form the antigen binding sites and costant regions that
anchor the receptors on the cell surface fig 16.12a.
The variable regions of the T cell receptors are encoded by multiple L-V, D, and J
gene segment

Major histocompatibility complex