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Cutting Edge: Developmental Stage-Specific Recruitment of Cohesin to CTCF Sites throughout Immunoglobulin Loci during B Lymphocyte Development¹

Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA 92037

Contraction of the large Igh and Ig loci brings all V genes, spanning >2.5 Mb in each locus, in proximity to DJ_H or J genes. CCCTC-binding factor (CTCF) is a transcription factor that regulates gene expression by long-range chromosomal looping. We therefore hypothesized that CTCF may be crucial for the contraction of the Ig loci, but no CTCF sites have been described in any V loci. Using ChIP-chip, we demonstrated many CTCF sites in the V_H and V regions. However, CTCF enrichment in the Igh locus, but not the Ig locus, was largely unchanged throughout differentiation, suggesting that CTCF binding alone cannot be responsible for stage-specific looping. Because cohesin can colocalize with CTCF, we performed chromatin immunoprecipitation for the cohesin subunit Rad21 and found lineage and stage-specific Rad21 recruitment to CTCF in all Ig loci. The differential binding of cohesin to CTCF sites may promote multiple loop formation and thus effective V(D)J recombination.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by National Institutes of Health Grants R01 A129672 and R01 A152313, S.C.D. was supported by National Institutes of Health Training Grant T32 HL07195.

² Address correspondence and reprint requests to Dr. Ann J. Feeney, Department of Immunology and Microbial Science, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037. E-mail address: feeney{at}scripps.edu

³ Abbreviations used in this paper: CTCF, CCCTC-binding factor; ChIP, chromatin immunoprecipitation; MEF, mouse embryonic fibroblast; qPCR, quantitative real-time PCR; 3'RR, 3' regulatory region.

⁴ The online version of this article contains supplemental material.

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