RAD51C interacts with RAD51B and is central to a larger protein complex in vivo exclusive of RAD51.
RAD51B and RAD51C are two of five known paralogs of the human RAD51 protein that are thought to function in both homologous recombination and DNA double-strand break repair. This work describes the in vitro and in vivo identification of the RAD51B/RAD51C heterocomplex. The RAD51B/RAD51C heterocomplex was isolated and purified by immunoaffinity chromatography from insect cells co-expressing the recombinant proteins. Moreover, co-immunoprecipitation of the RAD51B and RAD51C proteins from HeLa, MCF10A, and MCF7 cells strongly suggests the existence of an endogenous RAD51B/RAD51C heterocomplex. We extended these observations to examine the interaction between the RAD51B/RAD51C complex and the other RAD51 paralogs. Immunoprecipitation using protein-specific antibodies showed that RAD51C is central to a single large protein complex and/or several smaller complexes with RAD51B, RAD51D, XRCC2, and XRCC3. However, our experiments showed no evidence for the inclusion of RAD51 within these complexes. Further analysis is required to elucidate the function of the RAD51B/RAD51C heterocomplex and its association with the other RAD51 paralogs in the processes of homologous recombination and DNA double-strand break repair.
Miller
Kristi A
KA
Lawrence Livermore National Laboratory, Livermore, California 94550, USA.
Yoshikawa
Daniel M
DM
McConnell
Ian R
IR
Clark
Robin
R
Schild
David
D
Albala
Joanna S
JS
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D002845:Chromatography; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007313:Insecta; D011233:Precipitin Tests; D011485:Protein Binding; D051135:Rad51 Recombinase; D011994:Recombinant Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C413002:RAD51B protein, human; C448305:RAD51C protein, human; D011994:Recombinant Proteins; C495153:RAD51 protein, human; D051135:Rad51 Recombinase