Transcriptional regulation in early B cell development
Martin Fuxa1 and Jane A Skok1,2
Transcription factors and signalling molecules are important for
both lineage commitment and lineage-specific regulation. The
B cell specification factor Pax5 plays a dual role in B lineage
commitment. Simultaneously, it potentiates and limits
lineage choice by activating genes that are required for the B
cell program while repressing lineage-inappropriate genes;
more than 100 of the latter have now been identified. In this
context, repression of the tyrosine kinase Flt3 has been shown
to be essential for B lineage commitment. Regulation of antigen
receptor recombination constitutes another level at which
lineage specificity is determined, and the identification of two
factors, E47 and FOXP1, which regulate the activity of the
recombinase enzymes in B lineage cells, provides insight into
the mechanisms that determine this. New information
regarding the control of ordered recombination and allelic
exclusion comes from studies of cis-acting elements within the
Ig loci.
Addresses
1
The Department of Immunology and Molecular Pathology, Division of
Infection and Immunity, University College London, London W1T 4JF,
United Kingdom
2
Department of Pathology, New York University School of Medicine, 550
1st Avenue, MSB 531, New York, NY 10016, USA
Corresponding author: Skok, Jane A (jane.skok@med.nyu.edu)
Current Opinion in Immunology 2007, 19:129–136
This review comes from a themed issue on
Lymphocyte development
Edited by James Hagman and Dietmar Kappes
Available online 9th February 2007
0952-7915/$ – see front matter
# 2006 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.coi.2007.02.002
Introduction
Hematopoietic development is a highly regulated multi-
step process in which pluripotent hematopoietic stem
cells (HSCs) differentiate through intermediate progeni-
tors to produce all mature cell types of the blood. Limita-
tion of lineage choice during development is largely
regulated by a combination of transcription factors and
signalling pathways. B lymphopoiesis is initiated by the
entry of progenitors into the B-cell lineage transcription
programme and the concomitant sequential rearrange-
ment of the immunoglobulin genes through V(D)J recom-
bination. In this review, we will discuss various aspects of
early B-cell development and will focus on some of
the most recent advances in transcriptional regulation
of B-cell specification and differentiation.
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Lineage commitment in early B-cell
development
An early step in hematopoiesis is the commitment of
multipotent progenitors to common myeloid progenitors
[1] or lymphoid-primed multipotent progenitors [2]. The
latter subsequently differentiate to the earliest lymphocyte
progenitors, which upregulate the lymphoid-specific
recombinase genes Rag1 and Rag2 and give rise to common
lymphoid progenitors (CLPs) that have their characteristic
B, T and natural killer cell developmental potential [3].
The generation of lymphoid progenitors and B lympho-
cytes depends on signalling through the c-Kit, Flt3 and
IL-7 receptors; in their absence, early B cell progenitors in
the bone marrow are severely affected [4,5]. In addition,
developmental control of early B lymphopoiesis is
exerted by a regulatory network of key transcription
factors that include PU.1 (an ets-family member), Ikaros,
Bcl11a (a zinc finger transcription factor), E2A (a helix-
loop-helix protein), EBF (early B cell factor) and Pax5
(Figure 1) [3,6]. Whereas PU.1 appears to be crucial for
both myeloid and lymphoid progenitors [7], Ikaros con-
trols the development of lymphoid progenitors [8]. In the
absence of Bcl11a, early B lymphopoiesis is blocked at
the CLP stage before the pre-pro-B stage [9]. E2A and
EBF are considered primary B-cell fate determinants and
co-ordinately activate the expression of B cell specific
genes. However, the mere activation of the B lymphocyte
transcription program is not sufficient to commit B cell
progenitors to the B lymphoid lineage in the absence of
the paired domain protein Pax5. Targeted deletion of
Pax5 arrests B-cell development at the pro-B cell stage,
and these Pax5/ cells have the capacity to self-renew
extensively and retain a broad developmental potential
similar to that of multipotent progenitors [10,11].
Role of Pax5 in potentiating and limiting
lineage choice
Pax5 fulfils a dual role in B lineage commitment by
activating B cell specific genes while simultaneously
repressing lineage-inappropriate genes [11]. Target genes
that are activated by Pax5 code for essential components
of the pre-B- and B-cell receptor (pre-BCR and BCR,
respectively) signalling pathways [3]. These include Ebf1
(see Update), the receptor signalling chain Iga (encoded
by mb-1) [10], the stimulatory co-receptors CD19 [12] and
CD21 [13], and the central adaptor protein BLNK [14].
Lineage choices are limited by Pax5 acting as a transcrip-
tional repressor. In this capacity, the targets include
Notch1 and c-fms, which encodes M-CSFR (macrophage
colony stimulating factor receptor), which are important
for the differentiation of progenitors into myeloid
Current Opinion in Immunology 2007, 19:129–136
130 Lymphocyte development

Figure 1

Early B-cell development in the bone marrow. A schematic diagram of early B lymphopoiesis, showing the successive differentiation stages and
the rearrangement status of IgH and IgL genes. DH–JH rearrangements are initiated in the earliest lymphocyte progenitors (ELPs) at a low level
and are completed as the cell progresses to the CLP and pre-pro-B cell (also referred to as CLP2) stage. VH–DJH recombination takes place in
pro-B cells, and successful rearrangement leads to expression of the Igm protein as part of the pre-BCR in large pre-B cells. Subsequent
rearrangement of the IgL locus in small pre-B cells results in the expression of the BCR (consisting of m heavy and k or l light chains) on
immature B cells (Imm. B). The approximate points of the developmental arrest in mice that have defective transcription factors PU.1, Ikaros,
Bcl11a, E2A, EBF, Pax5 and Foxp1 (black) or signalling components involved in signalling through c-Kit, Flt3 or IL-7R (grey) are indicated above.
E2A- and EBF-deficient progenitors in the bone marrow resemble pre-pro-B cells but lack all Ig rearrangements. Abbreviation: HSC,
haematopoietic stem cell.

lineages and T cells, respectively [11,15] (Figure 2). A
recent microarray screen that investigated the repressor
function of Pax5 identified more than 100 target genes
[16]. Grouped according to category, these included
secreted proteins, cell adhesion molecules, signal trans-
ducers and nuclear proteins that are expressed in ery-
throid, myeloid and/or T lymphoid cells. Continuous
repression by Pax5 is known to be required, because
reactivation of these repressed targets is observed follow-
ing conditional deletion of Pax5 in pro-B and mature B
cells. Surprisingly, even at the transition to the plasma cell
stage, when Pax5 is physiologically lost during terminal
differentiation, repressed genes are reactivated and con-
tribute to the plasma cell transcriptional program [16].
Self-renewal capacity and developmental plasticity are
shared characteristics of Pax5/ pro-B cells and early
haematopoietic progenitors. Hence, a key function of
Pax5 in B-lineage commitment is repression of genes
that are required for maintaining the undifferentiated
state. Evidence in favour of this hypothesis comes from
the recent observation that Pax5 binds to the promoter
region of Flt3 and represses transcription [17]. This
tyrosine kinase receptor is required for efficient formation
of the CLP. In contrast to wild-type pro-B cells, Pax5-
deficient progenitors were shown to express Flt3 and to
proliferate in a dose-dependent manner in response to
Flt3L. Furthermore, enforced expression of Flt3 in hae-
matopoietic cells impairs B lymphopoiesis. These data
contribute to our understanding of the process by which
cells commit to the B lymphoid lineage by demonstrating
that repression of Flt3 by Pax5 is an essential step in this
process. Pax5-mediated gene repression was furthermore
shown to be essential for normal homeostasis of the
hematopoietic system, because ectopic expression of
the Pax5-repressed chemokine gene Ccl3 in B cells results
in increased osteoclast formation and bone loss [16].
Immunoglobulin gene rearrangement
Although the initial stages of B lineage commitment
depend upon the correct functioning of signalling path-
ways and transcription factors, ongoing differentiation is
additionally dependent on successful rearrangement of
immunoglobulin (Ig) gene loci, and stage-specific assembly
of immunoglobulin heavy chain (IgH) and light (IgL)
chain genes defines each phase of B-cell development.
In essence, recombination involves pairing of compatible
conserved recombination signal sequences that flank the
V, D and J gene segments of the Ig and T-cell receptor
(Tcr) gene segments [18–20]. Formation of a synaptic
complex and subsequent introduction of double-stranded
DNA breaks requires the recombinase enzymes RAG1
and RAG2. These proteins are expressed from an early
stage in lymphoid development and are present in lym-
phoid progenitor cells as well as in B and T cells [18–20].
The function of antigen receptor rearrangement is to
generate sufficient diversity to enable recognition of
any antigen encountered while at the same time ensuring
that each lymphocyte expresses a receptor that has a
single specificity. As a common machinery is required
for recombination of variable (V), diversity (D) and joining

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 Transcriptional regulation in early B cell development Fuxa and Skok 131

Figure 2

Dual role of Pax5 at B lineage commitment. Pax5 activates B cell specific genes (e.g. CD19, CD22 and mb-1) and simultaneously represses
lineage-inappropriate genes at B cell commitment. The latter are essential for diverse cellular functions such as cell–cell communication,
adhesion and migration in T lymphoid, myeloid and erythroid cells as well as in multipotent progenitors.

(J) gene segments in all immune receptor loci, devel-
opmentally ordered changes in accessibility are crucial for
maintaining the integrity of this process [21]. Regulation
of accessibility is exerted at several levels to ensure
lineage specificity, sequential rearrangement of IgH
and IgL genes and individual gene segments, and allelic
exclusion.
Lineage specificity
Enrichment of IgH and Igk at the nuclear periphery has
been shown to be important in limiting Ig recombination to
the appropriate lineage [22,23]. In non-B lineage cells, IgH
and Igk are anchored with their 50 ends positioned closest to
the periphery and their 30 ends located more centrally
[24,25]. At the IgH locus, a region of 1 Mb at the 50
end has been shown to be required for colocalization with
the nuclear lamina. Silencing of genes at the nuclear
periphery might result from an exclusion of transcription
factors, enrichment of transcriptional repressors or a com-
bination of both. Although expression of B lineage tran-
scription factors has been implicated in movement towards
the centre of the nucleus [23,24], the factors involved in
maintaining the default position of these loci at the per-
iphery in their silent state have not been identified to date.
It should be noted that, although peripheral location
is clearly important for limiting accessibility in non-B
lineage cells, it might play a different role at later stages of
B-cell development. It is now known that the nuclear
periphery can consist of more than one subcompartment,
favouring opposing outcomes for transcriptional regula-
tion. In yeast, nuclear pore association has been shown to
optimize transcription of an inducible gene [26,27];
location of genes in these regions could have a similar
function in mammalian cells and might explain the find-
ing that Ig loci are relocated to this subcompartment in
mature B lineage cells in which these loci are being
actively transcribed [28] (JAS, unpublished).
Ordered rearrangement
Recombination begins in the B-cell lineage at the pro-B
cell stage with rearrangement of the IgH locus, whereas
recombination of IgL takes place at the later pre-B cell
stage. Ordered recombination is not limited to the loci but
extends to the gene segments so that DH segments are
first joined to JH segments of the IgH loci, which is
followed by VH–DJH recombination.
The transcription factors E2A and EBF control the initial
DH–JH rearrangement step by activating expression of the
Rag genes and promoting accessibility of the DH–JH
region within the IgH locus [29,30]. PAX5, by contrast,
induces large-scale contraction of the IgH locus; this is
essential for the second stage of VH–DJH recombination,

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132 Lymphocyte development
enabling synapse formation of distant gene segments that
facilitates rearrangement [24]. RAG-mediated recombi-
nation is primarily determined by localized changes in
chromatin structure, which are regulated at multiple
levels including onset of sense and antisense germline
transcription [21,31], alterations in histone modifications
[32,33] and nucleosome remodelling [34].
Transcriptional control of the Rag genes themselves
constitutes another aspect of regulation. The previously
identified enhancer of RAG transcription (Erag) regulates
RAG expression specifically in B cells but not in T
lymphocytes, and its targeted deletion leads to a partial
block at the pro-B cell stage [35]. More insight into
transcriptional regulation of RAG expression comes from
two recent studies that demonstrate B lineage specific
control [36,37].
Borghesi et al. [36] show that loss of the transcription
factor E47, an E2A gene product, results in diminished
CLP and pre-pro-B subsets as well as a complete absence
of pro-B cells. Developmental progression to other
lineages is reduced but remains intact. In addition, this
study clearly demonstrates that E47 is required for RAG1
expression and recombinase activity as early as the CLP
stage. Interestingly, reduced recombinase expression is
apparent in B cells, occurs to a lesser extent in dendritic
and natural killer cells, but is absent in T cells.
In agreement with the B lineage specific requirements for
V(D)J recombination activity, Hu et al. [37] demonstrate
a significant role for the forkhead protein FOXP1 in this
context. Targeted deletion of Foxp1 results in embryonic
lethality, but reconstitution experiments using Foxp1/
fetal liver cells in Rag2-deficient recipients leads to a
dramatic reduction in mature B cell numbers and a differ-
entiation defect in the transition from pro-B to pre-B cells.
No obvious equivalent impairment in thymocyte devel-
opment was detected. Molecular characterization of these
Foxp1/ pro-B cells demonstrated reduced expression of
the B-cell regulator genes Tcfe2a (which encodes E2A) and
Ebf in addition to reduced expression of B cell specific
genes regulated downstream from the E2A and EBF path-
ways. These include Pax5, components of the pre-BCR,
and Rag1 and Rag2. Further analysis demonstrated that
Foxp1 mutant pro-B cells have a substantial reduction in
the frequency of VH–DJH recombination involving both
proximal and distal VH gene segments, whereas DH–JH
recombination is less severely affected. This phenotype is
reminiscent of the V(D)J recombination defect seen in
Erag/ mice [35]. The fact that both E2A and FOXP1
bind the Erag enhancer in vivo and exhibit similar loss-of-
function phenotypes suggests that these two transcription
factors and this cis-acting element in the RAG locus are
essential for regulating V(D)J recombinase activity during
B lymphopoiesis. The way in which they co-operate
remains to be determined.
Current Opinion in Immunology 2007, 19:129–136
Allelic exclusion
Allelic exclusion ensures monoallelic expression of anti-
gen receptor loci, which is essential for single specificity
antigen recognition. Accumulating evidence supports a
‘feedback inhibition’ model for establishing allelic
exclusion [38]. Functional rearrangement leads to
expression of Igm as part of the pre-BCR; this acts as
an important checkpoint preventing ongoing rearrange-
ment of the second DJH rearranged allele at the pre-B
cell stage of development when IgL rearrangement takes
place. Following a functional rearrangement at the IgH
locus, RAG protein expression is transiently downregu-
lated upon pre-BCR signalling. Signalling through the
IL-7R is attenuated at this stage and contributes to
allelic exclusion by decreasing accessibility at the IgH
locus [39]. Decontraction occurs at the IgH locus follow-
ing recombination and is instrumental in establishing
allelic exclusion as this process physically separates
distant gene segments, preventing further interaction.
In addition, following successful recombination of one
IgH allele, the second allele is repositioned at centro-
meric heterochromatin at the pre-B cell stage of devel-
opment while IgL recombination takes place [25].
Recruitment of the second allele to a repressive com-
partment of the nucleus is thought to act as a back-up
mechanism for establishing allelic exclusion by prevent-
ing accessibility to recombinase enzymes during IgL
recombination when the recombinase enzymes are once
again upregulated (Figure 3).
In contrast to the IgH locus, repositioning of the Igk locus
to centromeric clusters occurs at the pre-B cell stage of
development, limiting accessibility to a single allele
during recombination [25,40]. Although recruitment
of the IgH locus to centromeric clusters and deacetylation
have been correlated with a decrease in IL-7R signalling,
little is known about the mechanisms that govern regu-
lation of reversible locus contraction and repositioning of
loci to heterochromatic regions. Recently, however, a cis-
acting silencing element has been identified that has a
role in directing the Igk locus to centromeric heterochro-
matin and association with the transcription factor Ikaros
[41]. This silencer, known as Sis (silencer in the inter-
vening sequence), is located between the most 30 Vk gene
and the Jk gene segments and is the first cis-acting
element to be identified that is involved in centromeric
recruitment of the Ig loci.
Monoallelic silencing [40] and a low level of monoallelic
activation [42] have been implicated in allelic exclusion at
the Igk locus and, in this context, enhancer elements
might be important. Both IgH and Igk loci contain an
intronic enhancer located between the J gene segments
and the constant region. A recent study demonstrated that
deletion of the Igk locus intronic enhancer/matrix attach-
ment region (MiEk/) severely reduces accessibility and
rearrangement of this locus in pre-B cells [43]. This has
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 Transcriptional regulation in early B cell development Fuxa and Skok 133

Figure 3

Conformational changes and subnuclear relocation of immunoglobulin loci during B-cell development. (a) Germline organization of the murine Ig loci.
The mouse IgH and Igk loci together with the relative position of V gene segment families (yellow rectangles), indicating the most distal and proximal
family. In addition, D (orange vertical lines) and J segments (green vertical lines) as well as the constant regions (C, light blue rectangles) are shown.
Previously characterized enhancers and promotors are represented as red ovals and grey rectangles, respectively. The 1 Mb region that encompasses
the majority of the most distal VH gene family (J558) and appears to be important for adherence of the IgH locus to the nuclear periphery is depicted as
a dark grey bar. The location of the cis-acting element, Sis, which is required for targeting the Igk locus to centromeric heterochromatin, is shown as a
dark blue oval. The asterisk indicates the nonfunctional Jk pseudogene segment. (b) Subnuclear relocation and locus contraction of the IgH and Igk
loci. A diagram of the nucleus illustrates the location of the IgH and Igk loci and the rearrangement status at different stages of B-cell development. The
expression of Pax5, the pre-BCR and the BCR at the corresponding differentiation stages is indicated. The distal (50 ) and proximal (30 ) domains (dark
and light coloured circles, respectively) are depicted together with their location relative to the repressive compartments at the nuclear periphery (grey
shaded area) and centromeric heterochromatin (dark grey). The locus contraction/decontraction and monoallelic, centromeric recruitment states are
schematically shown for progenitors, pro-B, pre-B and activated B cells.

been attributed to reduced accessibility and germline
transcription. Replacing the endogenous intronic enhan-
cer of the Igk locus (MiEk) with the intronic heavy chain
enhancer (Em) promotes changes in accessibility and
greatly increases premature recombination of the Igk
locus at the pro-B cell stage of development. In addition,
Igk rearrangement is reduced at the pre-B cell stage. In
these EmR replacement loci, Igk rearrangement mimics
the normal stage-specific order of IgH recombination,
indicating a role for the Em enhancer in directing ordered
rearrangement. Further analysis will be required to deter-
mine whether these EmR replacement loci undergo pre-
mature changes in nuclear organization [25]. In addition,
the effect, if any, of enhancer replacement on IgH re-
arrangement remains unclear.
Conclusions
In this review, we have discussed recent advances in
transcriptional regulation of early B-cell development.
Detailed molecular analyses have provided important
insight into the gene repression function of Pax5 during
B-cell development. The transcriptional regulation of this
lineage commitment factor is determined by its inter-
action with distinct partner proteins. For instance, Pax5
cooperates with PU.1 to recruit co-repressors of the
Groucho protein family in a context-depending manner

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134 Lymphocyte development
to repressed target genes [3]. By contrast, cooperative
binding of Pax5 and Ets proteins to the mb-1 promoter
results in transcriptional activation [44]. Further studies
are necessary to identify other Pax5 interaction partners
that are required for the downregulation of diverse line-
age-inappropriate cellular functions and the activation of
the B-cell program during B-cell development.
Identification of E2A and FOXP1 as two key factors that
regulate recombinase activity specifically in B cells pro-
vides insight into the mechanisms that control lineage-
specific recombination. Although these proteins bind to a
previously characterized enhancer region in the Rag locus,
additional regulatory elements appear to be necessary for
lineage specificity. Further investigations will determine
the mechanism of transcription factor cooperation at these
sites.
Transcriptional regulators that influence changes in
nuclear location of Ig loci during B-cell development
remain to be identified. The molecular mechanisms that
underlie locus contraction are currently unknown and we
have little insight into the factors that regulate centro-
meric recruitment. Future investigations should aid in
characterizing the transcription factor composition of
distinct nuclear subcompartments, which will further
our understanding of these events.
Update
A recent study [45] that has analyzed differential regu-
lation of two promoters within the Ebf1 gene adds yet
another layer of complexity to the control of transcrip-
tional regulation during B lymphopoiesis. Activation of
the distal Ebf1 promoter is induced by E2A and EBF1 in
addition to IL-7 signaling, whereas the stronger proximal
promoter is activated by the combination of PAX5, ETS1
and PU.1. Absence of PAX5 impairs the function of the
proximal Ebf1 promoter and accumulation of EBF1
protein. In concert with this, the replication timing and
subnuclear localization of the Ebf1 locus are altered. This
indicates that the presence of an additional promoter can
further refine control of the expression of a locus and has
wider implications for feedback loops within the regulat-
ory network that orchestrates B cell development.
Acknowledgements
We thank Meinrad Busslinger and Adrian Erlebacher for critical reading of
the manuscript. This work was supported by a Wellcome trust project grant
and New York University School of Medicine start-up laboratory funding.
References and recommended reading
Papers of particular interest, published within the period of review,
have been highlighted as:
 of special interest
 of outstanding interest
1. Akashi K, Traver D, Miyamoto T, Weissman IL: A clonogenic
common myeloid progenitor that gives rise to all myeloid
lineages. Nature 2000, 404:193-197.
Current Opinion in Immunology 2007, 19:129–136
2. Adolfsson J, Mansson R, Buza-Vidas N, Hultquist A, Liuba K,
Jensen CT, Bryder D, Yang L, Borge OJ, Thoren LA et al.:
Identification of Flt3+ lympho-myeloid stem cells lacking
erythro-megakaryocytic potential a revised road map for adult
blood lineage commitment. Cell 2005, 121:295-306.
3. Busslinger M: Transcriptional control of early B cell
development. Annu Rev Immunol 2004, 22:55-79.
4. Waskow C, Paul S, Haller C, Gassmann M, Rodewald HR: Viable
c-KitW/W mutants reveal pivotal role for c-kit in the
maintenance of lymphopoiesis. Immunity 2002, 17:277-288.
5. Vosshenrich CA, Cumano A, Muller W, Di Santo JP, Vieira P:
Thymic stromal-derived lymphopoietin distinguishes fetal
from adult B cell development. Nat Immunol 2003, 4:773-779.
6. Medina KL, Singh H: Genetic networks that regulate B
lymphopoiesis. Curr Opin Hematol 2005, 12:203-209.
7. Dakic A, Metcalf D, Di Rago L, Mifsud S, Wu L, Nutt SL: PU.1
regulates the commitment of adult hematopoietic progenitors
and restricts granulopoiesis. J Exp Med 2005, 201:1487-1502.
8. Allman D, Miller JP: Common lymphoid progenitors, early
B-lineage precursors, and IL-7: characterizing the trophic and
instructive signals underlying early B cell development.
Immunol Res 2003, 27:131-140.
9. Liu P, Keller JR, Ortiz M, Tessarollo L, Rachel RA, Nakamura T,
Jenkins NA, Copeland NG: Bcl11a is essential for normal
lymphoid development. Nat Immunol 2003, 4:525-532.
10. Nutt SL, Urbanek P, Rolink A, Busslinger M: Essential functions
of Pax5 (BSAP) in pro-B cell development: difference between
fetal and adult B lymphopoiesis and reduced V-to-DJ
recombination at the IgH locus. Genes Dev 1997, 11:476-491.
11. Nutt SL, Heavey B, Rolink AG, Busslinger M: Commitment to the
B-lymphoid lineage depends on the transcription factor Pax5.
Nature 1999, 401:556-562.
12. Nutt SL, Morrison AM, Dorfler P, Rolink A, Busslinger M:
Identification of BSAP (Pax-5) target genes in early B-cell
development by loss- and gain-of-function experiments.
EMBO J 1998, 17:2319-2333.
13. Horcher M, Souabni A, Busslinger M: Pax5/BSAP maintains the
identity of B cells in late B lymphopoiesis. Immunity 2001,
14:779-790.
14. Schebesta M, Pfeffer PL, Busslinger M: Control of pre-BCR
signaling by Pax5-dependent activation of the BLNK gene.
Immunity 2002, 17:473-485.
15. Souabni A, Cobaleda C, Schebesta M, Busslinger M: Pax5
promotes B lymphopoiesis and blocks T cell development by
repressing Notch1. Immunity 2002, 17:781-793.
16. Delogu A, Schebesta A, Sun Q, Aschenbrenner K, Perlot T,
 Busslinger M: Gene repression by Pax5 in B cells is essential for
blood cell homeostasis and is reversed in plasma cells.
Immunity 2006, 24:269-281.
Identification of 110 Pax5-repressed genes provides important insight
into the repression function of Pax5 in B lineage commitment. These
genes fulfil diverse functions in different hematopoietic cell types and
encode proteins involved in receptor signalling, cell adhesion, migration,
transcriptional control and cellular metabolism. Expression analyses of
hematopoietic progenitors revealed that Pax5/ pro-B cells and com-
mon lymphoid progenitors display lymphoid and myeloid promiscuity of
gene expression. Conditional deletion of Pax5 in committed pro-B cells
and mature B cells demonstrates that Pax5 activity is continuously
required for the repression of these lineage-inappropriate genes.
Pax5-repressed genes are also reactivated in plasma cells, indicating
that the physiological loss of Pax5 during terminal differentiation con-
tributes to the plasma cell transcription program.
17. Holmes ML, Carotta S, Corcoran LM, Nutt SL: Repression of Flt3
 by Pax5 is crucial for B-cell lineage commitment.
Genes Dev 2006, 20:933-938.
The role of Pax5 in lineage commitment is extended by the finding that
Pax5 represses a crucial growth factor receptor, Flt3, which is required for
efficient formation of early lymphoid progenitors. Pax5-deficient pro-B
cells express abundant Flt3 that is rapidly silenced upon the introduction
of Pax5, which binds to the Flt3 promoter in vivo. Enforced expression of
Flt3 in wild-type progenitors significantly impairs B-cell development and
www.sciencedirect.com
 Transcriptional regulation in early B cell development Fuxa and Skok 135
indicates that the repression of Flt3 by Pax5 is essential for normal
B-lymphopoiesis.
18. Hesslein DG, Schatz DG: Factors and forces controlling V(D)J
recombination. Adv Immunol 2001, 78:169-232.
19. Bassing CH, Swat W, Alt FW: The mechanism and regulation
of chromosomal V(D)J recombination. Cell 2002,
109:S45-S55.
20. Schlissel MS: Regulating antigen-receptor gene assembly.
Nat Rev Immunol 2003, 3:890-899.
21. Yancopoulos GD, Alt FW: Developmentally controlled and
tissue-specific expression of unrearranged VH gene
segments. Cell 1985, 40:271-281.
22. Kosak ST, Skok JA, Medina KL, Riblet R, Le Beau MM, Fisher AG,
Singh H: Subnuclear compartmentalization of immunoglobulin
loci during lymphocyte development. Science 2002,
296:158-162.
23. Sato H, Saito-Ohara F, Inazawa J, Kudo A: Pax-5 is essential for k
sterile transcription during Ig k chain gene rearrangement.
J Immunol 2004, 172:4858-4865.
24. Fuxa M, Skok J, Souabni A, Salvagiotto G, Roldan E, Busslinger M:
Pax5 induces V-to-DJ rearrangements and locus contraction
of the immunoglobulin heavy-chain gene. Genes Dev 2004,
18:411-422.
25. Roldan E, Fuxa M, Chong W, Martinez D, Novatchkova M,
 Busslinger M, Skok JA: Locus ‘decontraction’ and centromeric
recruitment contribute to allelic exclusion of the
immunoglobulin heavy-chain gene. Nat Immunol 2005,
6:31-41.
This study uses three-dimensional DNA fluorescence in situ hybridiza-
tion analysis to demonstrate that distal V gene segments are brought
into contact with the proximal domains of Ig loci by DNA looping.
Reversible locus contraction facilitates synapse formation and recom-
bination of gene segments separated by a large distance in cells
that are undergoing recombination. Pre-BCR signalling induces
decontraction, which prevents ongoing recombination at the second
DJH rearranged IgH allele, and centromeric recruitment, which reposi-
tions the non-functional allele within a repressive subcompartment
of the nucleus during IgL rearrangement. Thus, both decontraction
and centromeric recruitment are important for establishing allelic
exclusion.
26. Taddei A, Gasser SM: Multiple pathways for telomere tethering:
functional implications of subnuclear position for
heterochromatin formation. Biochim Biophys Acta 2004,
1677:120-128.
27. Schmid M, Arib G, Laemmli C, Nishikawa J, Durussel T,
Laemmli UK: Nup-PI: the nucleopore-promoter interaction of
genes in yeast. Mol Cell 2006, 21:379-391.
28. Yang Q, Riblet R, Schildkraut CL: Sites that direct nuclear
compartmentalization are near the 50 end of the mouse
immunoglobulin heavy-chain locus. Mol Cell Biol 2005,
25:6021-6030.
29. Romanow WJ, Langerak AW, Goebel P, Wolvers-Tettero IL,
van Dongen JJ, Feeney AJ, Murre C: E2A and EBF act in synergy
with the V(D)J recombinase to generate a diverse
immunoglobulin repertoire in nonlymphoid cells. Mol Cell 2000,
5:343-353.
30. Goebel P, Janney N, Valenzuela JR, Romanow WJ, Murre C,
Feeney AJ: Localized gene-specific induction of
accessibility to V(D)J recombination induced by E2A and
early B cell factor in nonlymphoid cells. J Exp Med 2001,
194:645-656.
31. Bolland DJ, Wood AL, Johnston CM, Bunting SF, Morgan G,
Chakalova L, Fraser PJ, Corcoran AE: Antisense intergenic
transcription in V(D)J recombination. Nat Immunol 2004,
5:630-637.
32. McBlane F, Boyes J: Stimulation of V(D)J recombination by
histone acetylation. Curr Biol 2000, 10:483-486.
33. Chowdhury D, Sen R: Stepwise activation of the
immunoglobulin m heavy chain gene locus. EMBO J 2001,
20:6394-6403.
www.sciencedirect.com
34. Maes J, O’Neill LP, Cavelier P, Turner BM, Rougeon F,
Goodhardt M: Chromatin remodeling at the Ig loci prior to V(D)J
recombination. J Immunol 2001, 167:866-874.
35. Hsu LY, Lauring J, Liang HE, Greenbaum S, Cado D, Zhuang Y,
Schlissel MS: A conserved transcriptional enhancer regulates
RAG gene expression in developing B cells. Immunity 2003,
19:105-117.
36. Borghesi L, Aites J, Nelson S, Lefterov P, James P,
 Gerstein R: E47 is required for V(D)J recombinase activity
in common lymphoid progenitors. J Exp Med 2005,
202:1669-1677.
E47, an alternative splice product of the transcription factor E2A, is shown
to be required for expression of both Rag1 and V(D)J recombinase activity
in multipotent hematopoietic and early B-cell progenitors. The loss of E47
results in a ten-fold reduction of the CLP subset that lacks D–JH rear-
rangements and a profound arrest before the pro-B cell stage. Other
downstream lymphoid progeny of CLPs are, however, still intact in these
mice, albeit at reduced numbers. Although recombinase activity is inhib-
ited in early B lineage precursors in E47-deficient animals, loss of either
E47 or its cis-acting target Erag has little effect on recombinase activity in
thymic T lineage precursors. This report defines a role for E47 in lineage
progression at the CLP stage and in regulating B cell specific recombi-
nase activity.
37. Hu H, Wang B, Borde M, Nardone J, Maika S, Allred L, Tucker PW,
 Rao A: Foxp1 is an essential transcriptional regulator of B cell
development. Nat Immunol 2006, 7:819-826.
The forkhead transcription factor Foxp1 is essential for early B-cell
development and leads to a differentiation defect in the transition from
the pro-B cell to the pre-B cell stage. Foxp1 deficiency is associated with
decreased expression of all B lineage genes and of Rag1 and Rag2 in
early B-cell progenitors in the fetal liver. Foxp1 binds to the Erag enhancer
and its loss leads to impaired DH–JH and VH–DJH rearrangement in a B-
cell lineage specific way. The loss-of-function phenotype of Foxp1
resembles that of E2A+/ EBF+/ compound heterozygous mice. This
report identifies Foxp1 as an essential participant in the transcriptional
regulatory network of B lymphopoiesis.
38. Jung D, Giallourakis C, Mostoslavsky R, Alt FW: Mechanism and
control of V(D)J recombination at the immunoglobulin heavy
chain locus. Annu Rev Immunol 2006, 24:541-570.
39. Chowdhury D, Sen R: Transient IL-7/IL-7R signaling provides a
mechanism for feedback inhibition of immunoglobulin heavy
chain gene rearrangements. Immunity 2003, 18:229-241.
40. Goldmit M, Ji Y, Skok J, Roldan E, Jung S, Cedar H, Bergman Y:
 Epigenetic ontogeny of the Igk locus during B cell
development. Nat Immunol 2005, 6:198-203.
This study analyzes the epigenetic changes that occur at the Igk locus
during B-cell development. The authors report that, in pre-B cells, one
allele is preferentially packaged into active chromatin whereas the other
allele is repositioned at centromereric heterochromatin and is associated
with HP1 and Ikaros.
41. Liu Z, Widlak P, Zou Y, Xiao F, Oh M, Li S, Chang MY, Shay JW,
 Garrard WT: A recombination silencer that specifies
heterochromatin positioning and Ikaros association in the
immunoglobulin k locus. Immunity 2006, 24:405-415.
Using yeast artificial chromosome-based single-copy isotransgenic
mice, these authors identified a cis-acting element residing in the V–J
intervening sequence. This element, termed Sis, negatively regulates
rearrangement in this locus and appears necessary for targeting germ-
line Igk transgenes to centromeric heterochromatin in pre-B and B cells.
Sis is furthermore shown to associate with Ikaros — a repressor protein
that also colocalizes with centromeric heterochromatin. The data sug-
gest that Sis participates in the monoallelic silencing aspect of allelic
exclusion regulation through targeting to centromeric heterochromatin
domains.
42. Liang HE, Hsu LY, Cado D, Schlissel MS: Variegated
transcriptional activation of the immunoglobulin k locus in
pre-B cells contributes to the allelic exclusion of light-chain
expression. Cell 2004, 118:19-29.
43. Inlay MA, Lin T, Gao HH, Xu Y: Critical roles of the
 immunoglobulin intronic enhancers in maintaining the
sequential rearrangement of IgH and Igk loci. J Exp Med 2006,
203:1721-1732.
To identify cis-acting elements that regulate ordered rearrangement, the
authors of this study replaced the endogenous matrix attachment
region/Igk intronic enhancer (MiEk) with its heavy chain counterpart
Current Opinion in Immunology 2007, 19:129–136
136 Lymphocyte development

(Em). Replacement substantially increases accessibility of both Vk and
Jk segments to the recombinase enzymes in pro-B cells and induces Igk
rearrangement in these cells. Replacement loci, however, do not sup-
port Igk rearrangement in pre-B cells. This suggests an important role for
Em and MiEk in stage-specific ordered rearrangement.
44. Maier H, Ostraat R, Gao H, Fields S, Shinton SA, Medina KL,
Ikawa T, Murre C, Singh H, Hardy RR et al.: Early B cell factor
cooperates with Runx1 and mediates epigenetic changes
associated with mb-1 transcription. Nat Immunol 2004,
5:1069-1077.
45. Roessler S, Györy I, Imhof S, Spivakov M, Williams RR, Busslinger
M, Fisher AG, Grosschedl R: Distinct promoters mediate the
regulation of Ebf1 gene expression by IL-7 and Pax5.
Mol Cell Biol 2006, in press.

Endeavour
The quarterly magazine for the history and
philosophy of science.

You can access Endeavour online on

ScienceDirect, where you’ll find book reviews,
editorial comment and a collection of beautifully
illustrated articles on the history of science.

Featuring:

Information revolution: William Chambers, the publishing pioneer by A. Fyfe

Does history count? by K. Anderson
Waking up to shell shock: psychiatry in the US military during World War II by H. Pols
Deserts on the sea floor: Edward Forbes and his azoic hypothesis for a lifeless deep ocean by T.R. Anderson and T. Rice
‘Higher, always higher’: technology, the military and aviation medicine during the age of the two world wars by C. Kehrt
Bully for Apatosaurus by P. Brinkman

Coming soon:

Environmentalism out of the Industrial Revolution by C. Macleod

Pandemic in print: the spread of influenza in the Fin de Siècle by J. Mussell
Earthquake theories in the early modern period by F. Willmoth
Science in fiction - attempts to make a science out of literary criticism by J. Adams
The birth of botanical Drosophila by S. Leonelli

And much, much more. . .

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