The Fanconi anemia pathway promotes DNA glycosylase-dependent excision of interstrand DNA crosslinks.
Fanconi anemia (FA) is a recessive cancer prone syndrome featuring bone marrow failure and hypersensitivity to DNA interstrand crosslinks (ICLs) and, to a milder extension, to ionizing radiation and oxidative stress. Recently, we reported that human oxidative DNA glycosylase, NEIL1 excises with high efficiency the unhooked crosslinked oligomer within three-stranded DNA repair intermediate induced by photoactivated psoralen exposure. Complete reconstitution of repair of the ICL within a three-stranded DNA structure shows that it is processed in the short-patch base excision repair (BER) pathway. To examine whether the DNA damage hypersensitivity in FA cells follows impaired BER activities, we measured DNA glycosylase and AP endonuclease activities in cell-free extracts from wild-type, FA, and FA-corrected cells. We showed that immortalized lymphoid cells of FA complementation Groups A, C, and D and from control cells from normal donors contain similar BER activities. Intriguingly, the cellular level of NEIL1 protein strongly depends on the intact FA pathway suggesting that the hypersensitivity of FA cells to ICLs may, at least in part, arise from downregulation or degradation of NEIL1. Consistent with this result, plasmid-based expression of the FLAG-tagged NEIL1 protein partially complements the hypersensitivity FA cells to the crosslinking agents exposures, suggesting that NEIL1 specifically complements impaired capability of FA cells to repair ICLs and oxidative DNA damage. These findings shed light to how the FA pathway may regulate DNA repair proteins and bring explanation for the long-time disputed problem of the oxidative stress sensitive phenotype of FA cells.
Macé-Aimé
Gaëtane
G
CNRS UMR8200 Groupe, Voie FANC/BRCA et Cancer, Université Paris-Sud, Institut de Cancérologie Gustave Roussy, F-94805 Villejuif Cedex, France.
Couvé
Sophie
S
Khassenov
Bekbolat
B
Rosselli
Filippo
F
Saparbaev
Murat K
MK
D045744:Cell Line, Tumor; D003432:Cross-Linking Reagents / Q000494:pharmacology; D045647:DNA Glycosylases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D004260:DNA Repair; D015536:Down-Regulation; D005199:Fanconi Anemia / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015398:Signal Transduction
D003432:Cross-Linking Reagents; D045647:DNA Glycosylases; C473634:NEIL1 protein, human