Abstract
Genes encoding immunoglobulin heavy chains (Igh) are assembled by rearrangement of variable (VH), diversity (DH) and joining (JH) gene segments. Three critical constraints govern VH recombination. These include timing (VH recombination follows DH recombination), precision (VH gene segments recombine only to DJH junctions) and allele specificity (VH recombination is restricted to DJH-recombined alleles). Here we provide a model for these universal features of VH recombination. Analyses of DJH-recombined alleles showed that DJH junctions were selectively epigenetically marked, became nuclease sensitive and bound RAG recombinase proteins, which thereby permitted DH-associated recombination signal sequences to initiate the second step of Igh gene assembly. We propose that VH recombination is precise, because these changes did not extend to germline DH segments located 5′ of the DJH junction.
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Acknowledgements
Supported by the Intramural Research Program of the National Institute on Aging (US National Institutes of Health; AI20047 to F.W.A., and AI32524 to D.G.S.) and the Howard Hughes Medical Institute (F.W.A. and D.G.S.).
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R. Subrahmanyam designed and did all the experiments; R. Subrahmanyam and R. Sen analyzed and interpreted the data; H.D. assisted with Southern blot analysis; I.I. did the ChIP analysis of H3K4me2; T.C. assisted in the initial characterization of DJH-rearranged cell lines; Y.J. and D.G.S. provided D345 cells and discussions of the ChIP analysis of RAG; Y.Z. and F.W.A. generated the Eμ-deficient cell lines; R. Subrahmanyam and R. Sen wrote the manuscript; and D.G.S. and F.W.A. read and critiqued the manuscript.
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Subrahmanyam, R., Du, H., Ivanova, I. et al. Localized epigenetic changes induced by DH recombination restricts recombinase to DJH junctions. Nat Immunol 13, 1205–1212 (2012). https://doi.org/10.1038/ni.2447
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DOI: https://doi.org/10.1038/ni.2447
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