The Journal of Experimental Medicine
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Published online
doi:10.1084/jem.20071787
The Journal of Experimental Medicine, Vol. 204, No. 13, 3247-3256
The Rockefeller University Press, 0022-1007 $30.00
© Bates et al.
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ARTICLE

 Chromosomal position of a VH gene segment determines its activation and inactivation as a substrate for V(D)J recombination

Jamie Geier Bates, Dragana Cado, Hector Nolla, and Mark S. Schlissel

Division of Immunology, Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720

CORRESPONDENCE Mark S. Schlissel: mss{at}berkeley.edu

Complete IgHC gene rearrangement occurs only in B cells in a stage-specific and ordered manner. We used gene targeting to reposition a distal VH gene segment to a region just 5' of the DH gene cluster and found its activation to be highly dependent on the chromosomal domain within which it resides. The targeted VH gene segment rearranged at a higher frequency than its endogenous counterpart, its rearrangement was no longer ordered, and its ability to be silenced by allelic exclusion was lost. Additionally, the targeted VH gene segment lost lineage specificity, as VDJH rearrangement was observed in thymocytes. These data suggest that locus contraction, mimicked by proximal targeting, can override any regulation imposed by DNA sequences immediately surrounding VH gene segments.

Abbreviations used: BAC, bacterial artificial chromosome; cDNA, complementary DNA; ChIP, chromatin immunoprecipitation; DP, double positive; dsDNA, double-stranded DNA; hµ, human µ; icµ, intracellular µ; LM-PCR, ligation-mediated PCR; RSS, recombination signal sequence.

J.G. Bates's present address is Stanford University, Stanford, CA 94305.


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