Abstract

The overarching goal of this project for the past decade has been to elucidate the biochemical and molecular mechanisms of V(D)J recombination. We have been fortunate to discover a number of very interesting properties of the recombinase, and the new studies proposed here capitalize on a serendipitous convergence of investigations into mechanisms of cleavage and transposition. More exciting for us as immunologists, we are poised to bring this new molecular, mechanistic understanding to bear on large biological questions of autoimmunity and host defense. Specifically, recent work in my lab challenges the prevailing models of transposition and its role in oncogenic rearrangements, and suggests a new connection between repertoire restriction and autoimmunity In the three aims of this grant we propose to: I. Test a novel release and rebind model for RAG-mediated transposition; II. Investigate the effects of target structure and sequence on transposition and hybrid joint formation; III. Construct and analyze mice bearing RAG-1 allies with conditional recombination defects.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI036420-12
Application #
6925857
Study Section
Cellular and Molecular Immunology - B (CMI)
Program Officer
Kirkham, Perry M
Project Start
1994-08-01
Project End
2010-01-31
Budget Start
2005-02-15
Budget End
2006-01-31
Support Year
12
Fiscal Year
2005
Total Cost
$380,250
Indirect Cost

  Institution

Name
New York University
Department
Pathology
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016

  Publications

Bischerour, Julien; Lu, Catherine; Roth, David B et al. (2009) Base flipping in V(D)J recombination: insights into the mechanism of hairpin formation, the 12/23 rule, and the coordination of double-strand breaks. Mol Cell Biol 29:5889-99
Wong, Serre-Yu; Lu, Catherine P; Roth, David B (2008) A RAG1 mutation found in Omenn syndrome causes coding flank hypersensitivity: a novel mechanism for antigen receptor repertoire restriction. J Immunol 181:4124-30
Lu, Catherine P; Posey, Jennifer E; Roth, David B (2008) Understanding how the V(D)J recombinase catalyzes transesterification: distinctions between DNA cleavage and transposition. Nucleic Acids Res 36:2864-73
Lu, Catherine P; Sandoval, Hector; Brandt, Vicky L et al. (2006) Amino acid residues in Rag1 crucial for DNA hairpin formation. Nat Struct Mol Biol 13:1010-5
Posey, Jennifer E; Pytlos, Malgorzata J; Sinden, Richard R et al. (2006) Target DNA structure plays a critical role in RAG transposition. PLoS Biol 4:e350
Sandor, Zoltan; Calicchio, Monica L; Sargent, R Geoffrey et al. (2004) Distinct requirements for Ku in N nucleotide addition at V(D)J- and non-V(D)J-generated double-strand breaks. Nucleic Acids Res 32:1866-73
Lee, Gregory S; Neiditch, Matthew B; Salus, Sandra S et al. (2004) RAG proteins shepherd double-strand breaks to a specific pathway, suppressing error-prone repair, but RAG nicking initiates homologous recombination. Cell 117:171-84
Neiditch, Matthew B; Lee, Gregory S; Huye, Leslie E et al. (2002) The V(D)J recombinase efficiently cleaves and transposes signal joints. Mol Cell 9:871-8
Thai, To-Ha; Purugganan, Mary M; Roth, David B et al. (2002) Distinct and opposite diversifying activities of terminal transferase splice variants. Nat Immunol 3:457-62
Huye, Leslie E; Purugganan, Mary M; Jiang, Ming-Ming et al. (2002) Mutational analysis of all conserved basic amino acids in RAG-1 reveals catalytic, step arrest, and joining-deficient mutants in the V(D)J recombinase. Mol Cell Biol 22:3460-73

Showing the most recent 10 out of 35 publications