A kinetoplastid BRCA2 interacts with DNA replication protein CDC45.
The gene BRCA2, first identified as a breast cancer susceptibility locus in humans, encodes a protein involved in DNA repair in mammalian cells and mutations in this gene confer increased risk of breast cancer. Here we report a functional characterisation of a Trypanosoma brucei BRCA2 (TbBRCA2) orthologue and show that the protein interacts directly with TbRAD51. A further protein-protein interaction screen using TbBRCA2 identified other interacting proteins, including a trypanosome orthologue of CDC45 which is involved in initiation and progression of the replication fork complex during DNA synthesis. Deletion of the TbBRCA2 gene retards cell cycle progression during S-phase as judged by increased incorporation of BrdU and an increased percentage of cells with one nucleus and two kinetoplasts. These results provide insights into the potential role played by BRCA2 in DNA replication and reveal a novel interaction that couples replication and recombination in maintaining integrity of the genome.
Oyola
Samuel O
SO
Department of Pathology, University of Cambridge, UK. so515@york.ac.uk
Bringaud
Frédéric
F
Melville
Sara E
SE
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000502:physiology*; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000737:chemistry*; D004261:DNA Replication; D018105:DNA, Kinetoplast / Q000378:metabolism; D024522:Genes, BRCA2; D006801:Humans; D051135:Rad51 Recombinase / Q000737:chemistry* / Q000235:genetics; D014346:Trypanosoma brucei brucei / Q000378:metabolism*
C113571:CDC45 protein, human; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D018105:DNA, Kinetoplast; D051135:Rad51 Recombinase