The Journal of Experimental Medicine
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Published online 19 June 2006 doi:10.1084/jem.20052310
Rockefeller University Press, 0022-1007 $8.00
JEM, Volume 203, Number 7, 1721-1732
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ARTICLE

 Critical roles of the immunoglobulin intronic enhancers in maintaining the sequential rearrangement of IgH and Igk loci

Matthew A. Inlay, Tongxiang Lin, Heather H. Gao, and Yang Xu

Section of Molecular Biology, Division of Biological Sciences, University of California, San Diego, La Jolla, CA 92093

CORRESPONDENCE Yang Xu: yangxu{at}ucsd.edu

V(D)J recombination of immunoglobulin (Ig) heavy (IgH) and light chain genes occurs sequentially in the pro– and pre–B cells. To identify cis-elements that dictate this order of rearrangement, we replaced the endogenous matrix attachment region/Igk intronic enhancer (MiE{kappa}) with its heavy chain counterpart (Eµ) in mice. This replacement, denoted EµR, substantially increases the accessibility of both V{kappa} and J{kappa} loci to V(D)J recombinase in pro–B cells and induces Igk rearrangement in these cells. However, EµR does not support Igk rearrangement in pre–B cells. Similar to that in MiE{kappa}–/– pre–B cells, the accessibility of V{kappa} segments to V(D)J recombinase is considerably reduced in EµR pre–B cells when compared with wild-type pre–B cells. Therefore, Eµ and MiE{kappa} play developmental stage-specific roles in maintaining the sequential rearrangement of IgH and Igk loci by promoting the accessibility of V, D, and J loci to the V(D)J recombinase.

Abbreviations used: ChIP, chromatin immunoprecipitation; ES, embryonic stem; GT, germline transcript; hC{kappa}, human {kappa} constant region; LM, ligation mediated; MAR, matrix attachment region; mC{kappa}, mouse {kappa} constant region; MiE{kappa}, MAR/Igk intronic enhancer; MSRE-QPCR, methylation-sensitive restriction enzyme real-time quantitative PCR; PGK, phosphoglycerol kinase; RS, recombining sequence; RSS, recombination signal sequence; SE, signal end.

M.A. Inlay and T. Lin contributed equally to this work.


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