title: Multiple possible sites of BRCA2 interacting with DNA repair protein RAD51.
abstract: To investigate the biological consequences of aberrant BRCA2 protein during mammary carcinogenesis, we attempted to identify proteins that normally interact with BRCA2. By using a yeast two-hybrid system with a hybrid protein that contained residues 639-1,508 of BRCA2 protein fused to the GAL4 DNA-binding domain, we isolated five independent cDNA clones that encoded parts of RAD51 protein, a human homolog of bacterial RecA. In vitro experiments using anti-RAD51 antibody confirmed interaction of BRCA2 with RAD51. The RAD51-binding region of BRCA2 detected in the present study was distinct from the region reported recently. Further studies using smaller portions of BRCA2 defined at least two additional RAD51-binding domains, residues 982-1,066 and 1,139-1,266. Our results suggest that BRCA2 can interact with RAD51 through multiple sites of BRCA2 and that control of mitotic and meiotic recombination and/or of genomic integrity through binding to RAD51 may be a crucial mechanism by which BRCA2 suppresses abnormal proliferation of mammary cells.
authors.affiliation: Department of Human Genome Analysis, Japanese Foundation for Cancer Research, Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Katagiri
authors.forename: H
authors.initials: H
authors.lastname: Saito
authors.forename: A
authors.initials: A
authors.lastname: Shinohara
authors.forename: H
authors.initials: H
authors.lastname: Ogawa
authors.forename: N
authors.initials: N
authors.lastname: Kamada
authors.forename: Y
authors.initials: Y
authors.lastname: Nakamura
authors.forename: Y
authors.initials: Y
authors.lastname: Miki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D024682:BRCA2 Protein; D015153:Blotting, Western; D001943:Breast Neoplasms / Q000235:genetics*; D004254:DNA Nucleotidyltransferases / Q000235:genetics*; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D005982:Glutathione Transferase / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D051135:Rad51 Recombinase; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D024682:BRCA2 Protein; D004268:DNA-Binding Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D005982:Glutathione Transferase; D004254:DNA Nucleotidyltransferases; C495151:RAD51 protein, S cerevisiae; C495153:RAD51 protein, human; D051135:Rad51 Recombinase; C495154:Rad51 protein, mouse