We are interested in the safety mechanisms that prevent chromosome translocations that lead to the generation of lymphomas and leukemias, which are among the most common human cancers. With millions of lymphocytes undergoing V(D)J recombination each day, there are many opportunities for genomic integrity to be breached over the course of an individual's lifetime-- and chromosomal translocations are, in fact, a cardinal feature of lymphoid neoplasms. These rearrangements typically place an intact proto-oncogene under the regulatory control of the highly expressed Ig or TCR genes, leading to deleterious effects on cell growth, differentiation, or apoptosis. Compelling circumstantial evidence has implicated the V(D)J recombinase in these translocations, but how it might actually participate in aberrant rearrangements has remained mysterious. The Roth laboratory has recently made several discoveries that suggest novel mechanisms for V(D)J recombination-initiated aberrant rearrangements. We have also uncovered a new control point in the reaction that may prevent these events. The overarching goal of the studies proposed here is to understand the mechanisms that preserve genomic stability in lymphocytes and to define the molecular pathogenesis of V(D)J recombination-associated oncogenic rearrangements. To do so we will 1) probe the nature of the post-cleavage complex's scaffolding role; 2) examine the contribution of alternative RAG cleavage mechanisms to aberrant rearrangements; 3) create two lines of knock-in mice bearing RAG mutations we predict will increase the likelihood of aberrant rearrangements; 4) delineate the specific contributions of DNA damage sensors, checkpoint functions and the V(D)J recombinase to maintaining genomic stability. The last aim is in collaboration with the Petrini lab and capitalizes on their recent work implicating the Mrel I complex in controlling the fidelity of V(D)J recombination.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA104588-02
Application #
6879738
Study Section
Immunobiology Study Section (IMB)
Program Officer
Howcroft, Thomas K
Project Start
2004-04-01
Project End
2008-11-30
Budget Start
2005-04-01
Budget End
2006-03-31
Support Year
2
Fiscal Year
2005
Total Cost
$285,052
Indirect Cost
Name
New York University
Department
Pathology
Type
Schools of Medicine
DUNS #
121911077
City
New York
State
NY
Country
United States
Zip Code
10016
Hewitt, Susannah L; Wong, Jason B; Lee, Ji-Hoon et al. (2017) The Conserved ATM Kinase RAG2-S365 Phosphorylation Site Limits Cleavage Events in Individual Cells Independent of Any Repair Defect. Cell Rep 21:979-993
Mijuškovi?, Martina; Chou, Yi-Fan; Gigi, Vered et al. (2015) Off-Target V(D)J Recombination Drives Lymphomagenesis and Is Escalated by Loss of the Rag2 C Terminus. Cell Rep 12:1842-52
Gigi, Vered; Lewis, Susanna; Shestova, Olga et al. (2014) RAG2 mutants alter DSB repair pathway choice in vivo and illuminate the nature of 'alternative NHEJ'. Nucleic Acids Res 42:6352-64
Roth, David B (2014) V(D)J Recombination: Mechanism, Errors, and Fidelity. Microbiol Spectr 2:
Deriano, Ludovic; Roth, David B (2013) Modernizing the nonhomologous end-joining repertoire: alternative and classical NHEJ share the stage. Annu Rev Genet 47:433-55
Coussens, Marc A; Wendland, Rebecca L; Deriano, Ludovic et al. (2013) RAG2's acidic hinge restricts repair-pathway choice and promotes genomic stability. Cell Rep 4:870-8
Chaumeil, Julie; Micsinai, Mariann; Ntziachristos, Panagiotis et al. (2013) The RAG2 C-terminus and ATM protect genome integrity by controlling antigen receptor gene cleavage. Nat Commun 4:2231
Mijuskovic, Martina; Brown, Stuart M; Tang, Zuojian et al. (2012) A streamlined method for detecting structural variants in cancer genomes by short read paired-end sequencing. PLoS One 7:e48314
Deriano, Ludovic; Chaumeil, Julie; Coussens, Marc et al. (2011) The RAG2 C terminus suppresses genomic instability and lymphomagenesis. Nature 471:119-23
Arnal, Suzzette M; Holub, Abigail J; Salus, Sandra S et al. (2010) Non-consensus heptamer sequences destabilize the RAG post-cleavage complex, making ends available to alternative DNA repair pathways. Nucleic Acids Res 38:2944-54

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