The Organization and

Expression of Ig
Genes

Introduction
The vertebrate immune system is capable
of responding to an essentially infinite
array of foreign antigens
Variable vs. constant regions
Basis for variability organization and
expression of Ig genes

Overview
Historical perspective early theories
Multigene organization of Ig genes
Variable region gene rearrangements

of mechanism key players: DNA

signal sequences, specific enzymes

Basics

Generation of Ab diversity
seven

primary routes

Historical perspective

Ig sequence analysis revealed many

dilemmas
Extreme

diversity of Ab specificity
Variable regions vs constant regions
Isotypes with similar Ag specificity but
differing heavy-chain constant regions

Historical perspective

Proponents of the one-gene-one-protein

paradigm had trouble reconciling this model with
the oddities of Igs.
This led to an initial germ-line theory which
suggested that a significant portion of the
genome is dedicated solely to Ab coding.
Argument:

the immune system is THAT important

Historical perspective

In contrast, somatic-variation theories

emerged which suggested the opposite:
Relatively

small amount of Ig genes

Specificity arises from mutation and/or
recombination

Historical perspective

Dreyer & Bennet (1965)

Two-gene,

one-protein model
No precedent in any biological system

Historical perspective

Tonegawa & Hozumi (1976)

Compare

Ig DNA from embryonic (germline)

and adult myeloma (somatic) cells
Experimental data suggested that during
differentiation, the V and C genes undergo
rearrangement.
1987 Nobel prize

Tonegawa & Hozumi

Multigene Organization of Ig genes

Each class of Ig components (kappa,

lambda, heavy) encoded by separate
multigene families on different
chromosomes
Each

family contains several coding

sequences, or gene segments

Multigene Organization of Ig genes

Multigene Organization of Ig genes

& light chains: V (variable), J (joining), and C

(constant) gene segments
Heavy chains: V, D (diversity), J, and C gene
segments
A leader (L) sequence also precedes each V
segment.
Gene segments discovered by comparing DNA
sequences with amino acid sequences of Igs
Tonegawa,

again.

Organization of Ig germ-line
gene segments (mouse)

Pre-rearrangement!

Variable-region rearrangement

Multifaceted process, produces mature B

cells which are committed to express
specific Ab
Specificity

of Ab determined by the sequence

of its rearranged variable genes.

Light-chain rearrangements

V-J rearrangements
Specific

allowed rearrangements differ from

species to species, but a big-picture view
can suffice
Rearrangement occurs in ordered steps but
can be considered as random events which
result in the random determination of Ab
specificity

Kappa light-chain
rearrangement & RNA
processing

Leader sequence targets

nascent protein to ER
and is subsequently
cleaved

Heavy-chain rearrangements

Requires two separate rearrangement events

D-J

joining
V-DJ joining

Differential polyadenylation & RNA splicing can

result in mRNA with either Cu or C
heavy chain genes
B

cells can express BOTH IgM and IgD with identical

Ag specificity on its surface

Heavy-chain
rearrangement

IgM

IgD

The focus here is on

B cells

Mechanism of Variable region

DNA rearrangements

Recombination signal sequences direct recombination

Mechanism of Variable region

DNA rearrangements

Recombination signal sequences direct recombination

V, JK, VH, JH

VK, J, DH

Mechanism of Variable region

DNA rearrangements

A one-turn RSS can only join with a twoturn RSS

Why

might this be?

Mechanism of Variable region

DNA rearrangements
Gene segments are joined by a class of
enzymes called recombinases
Two recombination-activating genes
encode proteins which act together to
mediate V-(D)-J joining

RAG-1

RAG-2

a. Same
transcriptional
orientation
(most common)

Addn. of Pnucleotides
accomplished
with repair
enzymes

b. opposite
transcriptional
orientation

Mechanism of Variable region

DNA rearrangements

Rearrangements may be productive or

nonproductive.

Mechanism of Variable region

DNA rearrangements

Allelic exlusion
Heavy-chain

genes only expressed from one

chromosome
Light-chain genes only expressed from one
chromosome

Essential for specificity

Expression

of both alleles would result in a

multispecific B cell

Mechanism of Variable region

DNA rearrangements

Allelic exlusion

Generation of Ab diversity
Multiple germ-line gene segments
Combinatorial V-(D)-J joining
Junctional flexibility
P-region nucleotide addition
N-region nucleotide addition
Somatic hypermutation
Combinatorial association of light and
heavy chains

Possibly as high as 1010!

Junctional flexibility

P/N-addition

Somatic hypermutation

Nucleotide replacement, mediated by

activation-induced cytidine deaminase
(AID)
Also

plays a key role in class switching

Frequency of 10-3 per bp per generation

100,000X

the rate of spontaneous mutation!

Approx 1 mutation every 2 cell divisions

Somatic hypermutation