Abstract

Immune receptor gene rearrangement is central to the generation of antibody diversity. This somatic recombination event is mediated by conserved recombination signal sequences (RSS) which are found immediately adjacent to all functional immune receptor gene segments sequenced to date. Murine D(H) gene segments are flanked by equivalent signal sequences; however recombination via the V(H) proximal RSS appears to be precluded until the J(H) proximall RSS has been utilized. The proposed experiments should define both the sequence motif(s) and the trans-acting factors which are responsible for this phenomenon. Immune receptor rearrangement has been intensely studied for the last decade. Unfortunately, virtually nothing is known about the enzymology of this process. In 1989 and 1990, two highly conserved genes were isolated, """"""""recombinase activating genes 1 and 2"""""""" (RAG I and RAG 2), which are encoded within the same locus and which undoubtedly play a significant role in the somatic recombination of immune receptor genes. Though these genes were discovered nearly two years ago, biochemical information about these putative enzymes has yet to be reported. The baculovirus expresion system is being utilized to produce large amounts of these proteins so that biochemical, functional, and structural analyses can be performed. Ultimately, the goal of this research is to develop an in vitro recombination system to systematically study the enzymology of immune receptor rearrangement.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
1R01AI032600-01
Application #
3147733
Study Section
Allergy and Immunology Study Section (ALY)
Project Start
1992-03-01
Project End
1995-02-28
Budget Start
1992-03-01
Budget End
1993-02-28
Support Year
1
Fiscal Year
1992
Total Cost
Indirect Cost

  Institution

Name
University of Texas Sw Medical Center Dallas
Department
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390

  Publications

Reddy, Yeturu V R; Ding, Qi; Lees-Miller, Susan P et al. (2004) Non-homologous end joining requires that the DNA-PK complex undergo an autophosphorylation-dependent rearrangement at DNA ends. J Biol Chem 279:39408-13
Meek, Katheryn; Gupta, Shikha; Ramsden, Dale A et al. (2004) The DNA-dependent protein kinase: the director at the end. Immunol Rev 200:132-41
Yu, Yaping; Wang, Wei; Ding, Qi et al. (2003) DNA-PK phosphorylation sites in XRCC4 are not required for survival after radiation or for V(D)J recombination. DNA Repair (Amst) 2:1239-52
Ding, Qi; Reddy, Yeturu V R; Wang, Wei et al. (2003) Autophosphorylation of the catalytic subunit of the DNA-dependent protein kinase is required for efficient end processing during DNA double-strand break repair. Mol Cell Biol 23:5836-48
Kilzer, Jennifer M; Stracker, Travis; Beitzel, Brett et al. (2003) Roles of host cell factors in circularization of retroviral dna. Virology 314:460-7
Ding, Qi; Bramble, Lori; Yuzbasiyan-Gurkan, Vilma et al. (2002) DNA-PKcs mutations in dogs and horses: allele frequency and association with neoplasia. Gene 283:263-9
Woods, Timothy; Wang, Wei; Convery, Erin et al. (2002) A single amino acid substitution in DNA-PKcs explains the novel phenotype of the CHO mutant, XR-C2. Nucleic Acids Res 30:5120-8
Meek, K; Kienker, L; Dallas, C et al. (2001) SCID in Jack Russell terriers: a new animal model of DNA-PKcs deficiency. J Immunol 167:2142-50
Shin, E K; Rijkers, T; Pastink, A et al. (2000) Analyses of TCRB rearrangements substantiate a profound deficit in recombination signal sequence joining in SCID foals: implications for the role of DNA-dependent protein kinase in V(D)J recombination. J Immunol 164:1416-24
Kienker, L J; Shin, E K; Meek, K (2000) Both V(D)J recombination and radioresistance require DNA-PK kinase activity, though minimal levels suffice for V(D)J recombination. Nucleic Acids Res 28:2752-61

Showing the most recent 10 out of 17 publications