Abstract

Lymphocyte antigen receptor genes are assembled by a site-specific recombination process, referred to as V(D)J recombination, which is tightly regulated within the context of lymphocyte development. Furthermore, expression of antigen receptor chains is required for appropriate lymphocyte developmental stage progression. Therefore, understanding the mechanistic basis of V(D)J recombination will provide important insights into the molecular basis of lymphocyte development. T-cell receptor (TCR) beta gene rearrangement is ordered during development with D-beta to J-beta rearrangement preceding V-beta to D-beta rearrangement. Furthermore, V-beta to D-beta rearrangement is regulated in the context of allelic exclusion. They have shown that V-beta to D-beta rearrangement is mechanistically coupled through constraints placed on this step by the D-beta recombination signal sequence (RSS) beyond 12/23 compatibility. The goals of the studies described in this proposal are to elucidate the mechanisms responsible for coupling of V-beta to D-beta rearrangement by the 5' D-beta, RSS and to understand their role in regulating the ordered assembly of TCR beta genes during T-cell development. This will be accomplished through three specific aims. 1. Mutational analyses of the 5' D-beta RSS will be conducted to determine the regions responsible for coupling of V-beta to D-beta rearrangement. In addition, they will determine whether coupling by specific RSSs is generalizable to other rearrangement steps. 2. The mechanisms responsible for coupling V-beta to D-beta rearrangement by the 5' D-beta RSS will be elucidated. They will determine whether the requirement for the 5' D-beta RSS is strictly cis-acting. If trans-acting factors are required, they will attempt to isolate these factors. The role of transcriptional initiation, from the TATA box in the 5' D-beta RSS, in coupling V-beta to D-beta rearrangement will be determined. 3. They will determine whether TCR-beta allelic exclusion is accompanied by changes in transcriptional activity, methylation and/or chromatin structure of V-beta gene segments. In addition, the potential mechanistic role of V-beta to D-beta rearrangement coupling in effecting allelic exclusion will be assessed.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI047829-03
Application #
6532822
Study Section
Immunobiology Study Section (IMB)
Program Officer
Kirkham, Perry M
Project Start
2000-09-30
Project End
2005-05-31
Budget Start
2002-08-01
Budget End
2003-05-31
Support Year
3
Fiscal Year
2002
Total Cost
$277,200
Indirect Cost

  Institution

Name
Washington University
Department
Pathology
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130

  Publications

Fistonich, Chris; Zehentmeier, Sandra; Bednarski, Jeffrey J et al. (2018) Cell circuits between B cell progenitors and IL-7+ mesenchymal progenitor cells control B cell development. J Exp Med 215:2586-2599
Hung, Putzer J; Johnson, Britney; Chen, Bo-Ruei et al. (2018) MRI Is a DNA Damage Response Adaptor during Classical Non-homologous End Joining. Mol Cell 71:332-342.e8
Morales, Abigail J; Carrero, Javier A; Hung, Putzer J et al. (2017) A type I IFN-dependent DNA damage response regulates the genetic program and inflammasome activation in macrophages. Elife 6:
Bednarski, Jeffrey J; Pandey, Ruchi; Schulte, Emily et al. (2016) RAG-mediated DNA double-strand breaks activate a cell type-specific checkpoint to inhibit pre-B cell receptor signals. J Exp Med 213:209-23
Tubbs, Anthony T; Dorsett, Yair; Chan, Elizabeth et al. (2014) KAP-1 promotes resection of broken DNA ends not protected by ?-H2AX and 53BP1 in G?-phase lymphocytes. Mol Cell Biol 34:2811-21
KC, Wumesh; Satpathy, Ansuman T; Rapaport, Aaron S et al. (2014) L-Myc expression by dendritic cells is required for optimal T-cell priming. Nature 507:243-7
Dose, Marei; Emmanuel, Akinola Olumide; Chaumeil, Julie et al. (2014) ?-Catenin induces T-cell transformation by promoting genomic instability. Proc Natl Acad Sci U S A 111:391-6
Dorsett, Yair; Zhou, Yanjiao; Tubbs, Anthony T et al. (2014) HCoDES reveals chromosomal DNA end structures with single-nucleotide resolution. Mol Cell 56:808-18
Sandoval, Gabriel J; Graham, Daniel B; Bhattacharya, Deepta et al. (2013) Cutting edge: cell-autonomous control of IL-7 response revealed in a novel stage of precursor B cells. J Immunol 190:2485-9
Steinel, Natalie C; Lee, Baeck-Seung; Tubbs, Anthony T et al. (2013) The ataxia telangiectasia mutated kinase controls Ig? allelic exclusion by inhibiting secondary V?-to-J? rearrangements. J Exp Med 210:233-9

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