Deficiency in Incisions Produced by XPF at the Site of a DNA Interstrand Cross-Link in Fanconi Anemia Cells

• Published in: Biochemistry (Easton) 2007-12, Vol.46 (50), p.14359-14368
• Main Authors: Kumaresan, Kandallu R, Sridharan, Deepa M, McMahon, Laura W, Lambert, Muriel W
• Format: Article
• Language: English
• Subjects: Apoptosis; Base Sequence; Blotting, Western; Cell Line; DNA - chemistry; DNA - genetics; DNA - metabolism; DNA Damage; DNA Repair; DNA-Binding Proteins - metabolism; Electrophoresis, Polyacrylamide Gel; Fanconi Anemia - genetics; Fanconi Anemia - metabolism; Fanconi Anemia - pathology; Fanconi Anemia Complementation Group Proteins - genetics; Fanconi Anemia Complementation Group Proteins - metabolism; Humans; Molecular Sequence Data
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi7015958

Repair of DNA interstrand cross-links is a multistep process, critical to which is production of incisions at the site of the lesion resulting in the unhooking of the cross-link from DNA. We have previously shown that XPF is involved in production of incisions at the site of a psoralen interstrand cross-link and that in Fanconi anemia, complementation group A (FA-A) cells, there is a deficiency in these incisions. We now demonstrate that in FA complementation group B, C, D2, F, and G cells there is also a deficiency in production of these incisions. Involvement of FA proteins in this process is demonstrated by the ability of FA cells, corrected with the appropriate FANC cDNAs, to produce these incisions and by inhibition of these incisions by antibodies against these proteins. This incision deficiency correlates with reduced levels of DNA repair synthesis in these cells and is not due to reduced levels of XPF. FA proteins could be influencing this incision process by interacting either with proteins involved in the unhooking step or with damaged DNA, acting as a damage sensor. The results also demonstrate that FA cells are undergoing apoptosis by 12 h after interstrand cross-link damage. It is thus proposed that the single-strand breaks known to be created in DNA during apoptosis could mask the deficiency in ability of FA cells to incise cross-linked DNA and could account for the reported discrepancy as to whether FA cells are deficient in the incision step of the repair process.