Modulation of RAG/DNA complex by HSP70 in V(D)J recombination.
V(D)J recombination, a site-specific gene rearrangement process, requires two RAG1 and RAG2 proteins specifically recognizing recombination signal sequences and forming DNA double-strand breaks. The broken DNA ends tightly bound to RAG proteins are joined by repair proteins. Here, we found that heat shock protein 70 was associated with RAG2 following two-step affinity chromatography purification. It was also co-immunoprecipitated with RAG2 in pro-B cells. Purified HSP70 protein disrupted RAG/DNA complexes assembled in vitro and also inhibited the V(D)J cleavage (both nick and hairpin formation) in a dose-dependent manner. This HSP70 action required ATP energy. These data suggest that HSP70 might play a crucial role in disassembling RAG/DNA complexes stably formed during V(D)J recombination.
Son
Yong Mi
YM
Department of Biochemistry and MRCND and Institute of Health Sciences, Gyeongsang National University School of Medicine, 92 Chilam-dong, JinJu 660-751, Republic of Korea.
Lee
Jung Hwa
JH
Kim
Deok Ryong
DR
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D015322:Gene Rearrangement, B-Lymphocyte; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism*; D011995:Recombination, Genetic; D045567:VDJ Recombinases / Q000378:metabolism*
D004268:DNA-Binding Proteins; D018840:HSP70 Heat-Shock Proteins; D009687:Nuclear Proteins; C499311:RAG2 protein, human; D000255:Adenosine Triphosphate; D004247:DNA; D045567:VDJ Recombinases