Inducibility of nuclear Rad51 foci after DNA damage distinguishes all Fanconi anemia complementation groups from D1/BRCA2.
Fanconi anemia (FA) is a cancer susceptibility disorder characterized by chromosomal instability and hypersensitivity to DNA cross-linking agents. So far 11 complementation groups have been identified, from which only FA-D1/BRCA2 and FA-J are defective downstream of the central FANCD2 protein as cells from these groups are capable of monoubiquitinating FANCD2. In this study we show that cells derived from patients from the new complementation groups, FA-I, FA-J and FA-L are all proficient in DNA damage induced Rad51 foci formation, making the cells from FA-D1/BRCA2 patients that are defective in this process the sole exception. Although FA-B patient HSC230 was previously reported to also have biallelic BRCA2 mutations, we found normal Rad51 foci formation in cells from this patient, consistent with the recent identification of an X-linked gene being mutated in four unrelated FA-B patients. Thus, our data show that none of the FA proteins, except BRCA2, are required to sequester Rad51 into nuclear foci. Since cells from the FA-D1 and FA-J patient groups are both able to monoubiquitinate FANCD2, the "Rad51 foci phenotype" provides a convenient assay to distinguish between these two groups. Our results suggest that FANCJ and FANCD1/BRCA2 are part of the integrated FANC/BRCA DNA damage response pathway or, alternatively, that they represent sub-pathways in which only FANCD1/BRCA2 is directly connected to the process of homologous recombination.
Godthelp
Barbara C
BC
Department of Toxicogenetics, Leiden University Medical Center, Wassenaarseweg 72, NL-2333 AL Leiden, The Netherlands.
Wiegant
Wouter W
WW
Waisfisz
Quinten
Q
Medhurst
Annette L
AL
Arwert
Fré
F
Joenje
Hans
H
Zdzienicka
Małgorzata Z
MZ
D024682:BRCA2 Protein / Q000235:genetics* / Q000378:metabolism; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004249:DNA Damage / Q000235:genetics*; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism; D052240:Fanconi Anemia Complementation Group L Protein / Q000235:genetics*; D005347:Fibroblasts / Q000378:metabolism; D006801:Humans; D051135:Rad51 Recombinase / Q000096:biosynthesis* / Q000235:genetics*; D011995:Recombination, Genetic
D024682:BRCA2 Protein; D052240:Fanconi Anemia Complementation Group L Protein; C495153:RAD51 protein, human; D051135:Rad51 Recombinase