title: BRCA2 mediates centrosome cohesion via an interaction with cytoplasmic dynein.
abstract: BRCA2 is responsible for familial breast and ovarian cancer and has been linked to DNA repair and centrosome duplication. Here we analyzed the mechanism by which the centrosomal localization signal (CLS) of BRCA2 interacts with cytoplasmic dynein 1 to localize BRCA2 to the centrosome. In vitro pull-down assays demonstrated that BRCA2 directly binds to the cytoplasmic dynein 1 light intermediate chain 2. A dominant-negative HA-CLS-DsRed fusion protein, the depletion of dynein by siRNA, and the inactivation of dynein by EHNA, inhibited the localization of BRCA2 at centrosomes and caused the separation of centrosome pairs during the S-phase. The double depletion of BRCA2 and C-Nap1 caused a larger dispersion of centrosome distances than the silencing of C-Nap1. These results suggest that cytoplasmic dynein 1 binds to BRCA2 through the latter's CLS and BRCA2 mediates the cohesion between centrosomes during the S phase, potentially serving as a cell-cycle checkpoint.
authors.affiliation: a Department of Molecular Genetics , Medical Research Institute, Tokyo Medical and Dental University , Tokyo , Japan.
authors.forename: Sadiya
authors.initials: S
authors.lastname: Malik
authors.affiliation: b Department of Genetic Diagnosis , The Cancer Institute, Japanese Foundation for Cancer Research , Tokyo , Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Saito
authors.affiliation: a Department of Molecular Genetics , Medical Research Institute, Tokyo Medical and Dental University , Tokyo , Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Takaoka
authors.affiliation: a Department of Molecular Genetics , Medical Research Institute, Tokyo Medical and Dental University , Tokyo , Japan.|b Department of Genetic Diagnosis , The Cancer Institute, Japanese Foundation for Cancer Research , Tokyo , Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Miki
authors.affiliation: a Department of Molecular Genetics , Medical Research Institute, Tokyo Medical and Dental University , Tokyo , Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Nakanishi
mesh_terms: D000595:Amino Acid Sequence; D024682:BRCA2 Protein / Q000378:metabolism*; D018385:Centrosome / Q000378:metabolism*; D056445:Cytoplasmic Dyneins / Q000737:chemistry / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D008954:Models, Biological; D050505:Mutant Proteins / Q000378:metabolism; D011485:Protein Binding; D016196:S Phase
chemical_list: D024682:BRCA2 Protein; C551750:BRCA2 protein, human; D050505:Mutant Proteins; D056445:Cytoplasmic Dyneins
keywords: BRCA2; C-Nap1; CLS; LIC2; centrosome; dynein