title: Nuclear localization of Rad51B is independent of Rad51C and BRCA2.
abstract: Rad51B is one of the five paralogs of human Rad51 and is found in a multiprotein complex with three other Rad51 paralogs, Rad51C, Rad51D and Xrcc2. Participation of Rad51B in this complex depends on its direct interaction with Rad51C. Examination of EGFP-Rad51B fusion protein in HeLa S3 cells and immunofluorescence in several human cell lines reveal the nuclear localization of Rad51B. Mutations in the N-terminal KKLK motif of Rad51B (amino acids 4-7), result in the cytoplasmic localization of Rad51B suggesting that the KKLK sequence is the nuclear localization signal (NLS) for the Rad51B protein. Examination of wild-type EGFP-Rad51B fusion protein in hamster irs3 mutant cells, deficient in Rad51C, showed that Rad51B localizes to the nucleus independently of Rad51C, the only known direct binding partner for Rad51B. Utilization of a BRCA2 mutant cell line, CAPAN-1, showed that Rad51B also localizes to the nucleus independent of BRCA2. Although both Rad51B and BRCA2 are clearly involved in the homologous recombinational repair pathway, Rad51B and BRCA2 do not appear to associate. This study finds that a KKLK motif in the N-terminus of Rad51B serves as an NLS that allows Rad51B to localize to the nucleus independent of Rad51C or BRCA2.
authors.affiliation: Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, 7000 East Avenue, L-448, Livermore, CA 94550, USA.
authors.forename: Kristi A
authors.initials: KA
authors.lastname: Miller
authors.forename: John M
authors.initials: JM
authors.lastname: Hinz
authors.forename: N Alice
authors.initials: NA
authors.lastname: Yamada
authors.forename: Lawrence H
authors.initials: LH
authors.lastname: Thompson
authors.forename: Joanna S
authors.initials: JS
authors.lastname: Albala
mesh_terms: D024682:BRCA2 Protein / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006801:Humans
chemical_list: D024682:BRCA2 Protein; D004268:DNA-Binding Proteins; C413002:RAD51B protein, human; C448305:RAD51C protein, human