Xrcc3 is required for assembly of Rad51 complexes in vivo.
Rad51 is a member of a family of eukaryotic proteins related to the bacterial recombinational repair protein RecA. Rad51 protein localizes to multiple subnuclear foci in Chinese hamster ovary cells. Subnuclear Rad51 foci are induced by ionizing radiation or the DNA cross-linking agent cisplatin. Formation of these foci is likely to reflect assembly of a multimeric form of Rad51 that promotes DNA repair. Formation of damage-induced Rad51 foci does not occur in the Chinese hamster ovary cell line irs1SF, which is sensitive to DNA damaging agents. The Rad51 focus formation defect of irs1SF cells is corrected by a construct that encodes the repair protein Xrcc3. Xrcc3 is a human homolog of Rad51 previously isolated by virtue of its ability to correct the radiation sensitivity of irs1SF cells. Changes in the steady state level of Rad51 protein do not account for the irs1SF defect nor do they account for the appearance of foci following DNA damage. These results suggest that Xrcc3 is required for the assembly or stabilization of a multimeric form of Rad51 during DNA repair. Cell lines defective in two different components of DNA protein kinase formed Rad51 foci in response to damage, indicating DNA protein kinase is not required for damaged-induced mobilization of Rad51.
Bishop
D K
DK
Department of Radiation and Cellular Oncology, University of Chicago, Chicago, Illinois 60637, USA. dbishop@medway.uchicago.edu
Ear
U
U
Bhattacharyya
A
A
Calderone
C
C
Beckett
M
M
Weichselbaum
R R
RR
Shinohara
A
A
D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D016466:CHO Cells; D002467:Cell Nucleus / Q000528:radiation effects; D002945:Cisplatin / Q000494:pharmacology; D006224:Cricetinae; D004249:DNA Damage; D004260:DNA Repair; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D051135:Rad51 Recombinase; D011838:Radiation-Sensitizing Agents / Q000494:pharmacology
D000970:Antineoplastic Agents; D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D046911:Macromolecular Substances; D011838:Radiation-Sensitizing Agents; C110901:X-ray repair cross complementing protein 3; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; D002945:Cisplatin