title: Double-strand break repair-independent role for BRCA2 in blocking stalled replication fork degradation by MRE11.
abstract: Breast cancer suppressor BRCA2 is critical for maintenance of genomic integrity and resistance to agents that damage DNA or collapse replication forks, presumably through homology-directed repair of double-strand breaks (HDR). Using single-molecule DNA fiber analysis, we show here that nascent replication tracts created before fork stalling with hydroxyurea are degraded in the absence of BRCA2 but are stable in wild-type cells. BRCA2 mutational analysis reveals that a conserved C-terminal site involved in stabilizing RAD51 filaments, but not in loading RAD51 onto DNA, is essential for this fork protection but dispensable for HDR. RAD51 filament disruption in wild-type cells phenocopies BRCA2 deficiency. BRCA2 prevents chromosomal aberrations on replication stalling, which are alleviated by inhibition of MRE11, the nuclease responsible for this form of fork instability. Thus, BRCA2 prevents rather than repairs nucleolytic lesions at stalled replication forks to maintain genomic integrity and hence likely suppresses tumorigenesis through this replication-specific function.
authors.affiliation: Developmental Biology Program, Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA. schlachk@mskcc.org
authors.forename: Katharina
authors.initials: K
authors.lastname: Schlacher
authors.forename: Nicole
authors.initials: N
authors.lastname: Christ
authors.forename: Nicolas
authors.initials: N
authors.lastname: Siaud
authors.forename: Akinori
authors.initials: A
authors.lastname: Egashira
authors.forename: Hong
authors.initials: H
authors.lastname: Wu
authors.forename: Maria
authors.initials: M
authors.lastname: Jasin
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D024682:BRCA2 Protein / Q000378:metabolism*; D002460:Cell Line; D002470:Cell Survival; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000378:metabolism*; D042822:Genomic Instability; D006801:Humans; D000076228:MRE11 Homologue Protein; D008969:Molecular Sequence Data; D016415:Sequence Alignment
chemical_list: D024682:BRCA2 Protein; D004268:DNA-Binding Proteins; C121766:MRE11 protein, human; C448305:RAD51C protein, human; D000076228:MRE11 Homologue Protein