Direct interaction of the Fanconi anaemia protein FANCG with BRCA2/FANCD1.
Fanconi anaemia (FA) is an autosomal recessive genetic disorder characterized by progressive bone marrow failure, multiple congenital abnormalities, and an increased risk of cancer. FA cells are characterized by chromosomal instability and hypersensitivity to DNA interstrand crosslinking agents. At least eight complementation groups exist (FA-A to G), and the genes for all of these except FA-B have been cloned. Functional linkage between the FA pathway and genes involved in susceptibility to breast cancer has been demonstrated by the interaction of the FANCA and FANCD2 proteins with BRCA1, and the discovery that the FANCD1 gene is identical to BRCA2. Here we have used the yeast two-hybrid system to test for direct interaction between BRCA2 or its effector RAD51 and the FANCA, FANCC and FANCG proteins. We found that FANCG was capable of binding to two separate sites in the BRCA2 protein, located either side of the BRC repeats. Furthermore, FANCG could be co-immunoprecipitated with BRCA2 from human cells, and FANCG co-localized in nuclear foci with both BRCA2 and RAD51 following DNA damage with mitomycin C. These results demonstrate that BRCA2 is directly connected to a pathway that is deficient in interstrand crosslink repair, and that at least one other FA protein is closely associated with the homologous recombination DNA repair machinery.
Hussain
Shobbir
S
Division of Genetics and Development, Guy's, King's and St Thomas' School of Medicine, King's College London, UK.
Witt
Emily
E
Huber
Pia A J
PA
Medhurst
Annette L
AL
Ashworth
Alan
A
Mathew
Christopher G
CG
D000595:Amino Acid Sequence; D024682:BRCA2 Protein / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D018797:Cell Cycle Proteins; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004249:DNA Damage / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D052217:Fanconi Anemia Complementation Group A Protein; D052218:Fanconi Anemia Complementation Group C Protein; D052241:Fanconi Anemia Complementation Group G Protein; D051856:Fanconi Anemia Complementation Group Proteins; D005786:Gene Expression Regulation; D005808:Genes, Recessive; D017930:Genes, Reporter; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D016685:Mitomycin / Q000494:pharmacology; D008954:Models, Biological; D009687:Nuclear Proteins; D011233:Precipitin Tests; D011506:Proteins / Q000378:metabolism; D051135:Rad51 Recombinase; D020798:Two-Hybrid System Techniques
D024682:BRCA2 Protein; D018797:Cell Cycle Proteins; D003432:Cross-Linking Reagents; D004268:DNA-Binding Proteins; C498471:FANCA protein, human; C497651:FANCC protein, human; C499909:FANCG protein, human; D052217:Fanconi Anemia Complementation Group A Protein; D052218:Fanconi Anemia Complementation Group C Protein; D052241:Fanconi Anemia Complementation Group G Protein; D051856:Fanconi Anemia Complementation Group Proteins; D009687:Nuclear Proteins; D011506:Proteins; D016685:Mitomycin; C495153:RAD51 protein, human; D051135:Rad51 Recombinase