The genes which encode immunoglobulin and T cell receptor molecules are assembled from V, D, and J gene-segments by a novel series of site-specific DNA recombination reactions known as V(D)J recombination. The recombinase consists of the lymphocyte-specific proteins RAG1 and RAG2 and the widely expressed non-homologous end-joining proteins. The recombinase recognizes conserved recombination signal sequences (RSSs) which flank gene segments and introduces dsDNA breaks which are repaired to form coding and signal joints. Recombination at the seven complex antigen-receptor loci in B and T cells is regulated with respect to lineage specificity, order within a lineage, and allelic exclusion. We and others have shown that V(D)J recombination is regulated in part by the accessibility of RSSs within chromatin structure and that germline transcription of unrearranged gene segments either reflects or actually causes chromatin accessibility. The central goal of this proposal is to understand, at the biochemical level, the mechanisms which dictate the biological regulation of V(D)J recombination. This goal will be pursued through pursuit of the following specific aims: 1) to determine the location and structure of nucleosomes across the J-kappa cluster of gene segments and how nucleosomes are disrupted to allow recombinase accessibility; 2) to determine whether the V(D)J recombinase nicks DNA in vivo and whether the conversion from nicked to broken DNA is a regulated step in the reaction; 3) to determine the catalytic properties in vivo of a core-domain mutant of RAG2; and 4) to define the role of transcriptional regulatory elements in the regulation of V(D)J recombination. This work is significant because of the essential role of antigen receptors in acquired immunity and the potential involvement of the recombinase in generation of chromosomal translocations associated with lymphoid malignancies. ? ? ? ?

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Method to Extend Research in Time (MERIT) Award (R37)
Project #
Application #
Study Section
Cellular and Molecular Immunology - B (CMI)
Program Officer
Kraemer, Kristy A
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
University of California Berkeley
Schools of Arts and Sciences
United States
Zip Code
Vettermann, Christian; Timblin, Greg A; Lim, Vivian et al. (2015) The proximal J kappa germline-transcript promoter facilitates receptor editing through control of ordered recombination. PLoS One 10:e0113824
Chow, Kwan T; Schulz, Danae; McWhirter, Sarah M et al. (2013) Gfi1 and gfi1b repress rag transcription in plasmacytoid dendritic cells in vitro. PLoS One 8:e75891
Timblin, Greg A; Schlissel, Mark S (2013) Ebf1 and c-Myb repress rag transcription downstream of Stat5 during early B cell development. J Immunol 191:4676-87
Degner, Stephanie C; Verma-Gaur, Jiyoti; Wong, Timothy P et al. (2011) CCCTC-binding factor (CTCF) and cohesin influence the genomic architecture of the Igh locus and antisense transcription in pro-B cells. Proc Natl Acad Sci U S A 108:9566-71
Guo, Chunguang; Yoon, Hye Suk; Franklin, Andrew et al. (2011) CTCF-binding elements mediate control of V(D)J recombination. Nature 477:424-30
Sukumar, Selvakumar; Schlissel, Mark S (2011) Receptor editing as a mechanism of B cell tolerance. J Immunol 186:1301-2
Vettermann, Christian; Schlissel, Mark S (2010) Allelic exclusion of immunoglobulin genes: models and mechanisms. Immunol Rev 237:22-42
Hewitt, Susannah L; Farmer, Deborah; Marszalek, Katarzyna et al. (2008) Association between the Igk and Igh immunoglobulin loci mediated by the 3'Igk enhancer induces 'decontraction'of the Igh locus in pre-B cells. Nat Immunol 9:396-404
Bates, Jamie Geier; Cado, Dragana; Nolla, Hector et al. (2007) Chromosomal position of a VH gene segment determines its activation and inactivation as a substrate for V(D)J recombination. J Exp Med 204:3247-56
Curry, John D; Schulz, Danae; Guidos, Cynthia J et al. (2007) Chromosomal reinsertion of broken RSS ends during T cell development. J Exp Med 204:2293-303

Showing the most recent 10 out of 18 publications