ATR-dependent phosphorylation of FANCA on serine 1449 after DNA damage is important for FA pathway function.
Previous work has shown several proteins defective in Fanconi anemia (FA) are phosphorylated in a functionally critical manner. FANCA is phosphorylated after DNA damage and localized to chromatin, but the site and significance of this phosphorylation are unknown. Mass spectrometry of FANCA revealed one phosphopeptide, phosphorylated on serine 1449. Serine 1449 phosphorylation was induced after DNA damage but not during S phase, in contrast to other posttranslational modifications of FA proteins. Furthermore, the S1449A mutant failed to completely correct a variety of FA-associated phenotypes. The DNA damage response is coordinated by phosphorylation events initiated by apical kinases ATM (ataxia telangectasia mutated) and ATR (ATM and Rad3-related), and ATR is essential for proper FA pathway function. Serine 1449 is in a consensus ATM/ATR site, phosphorylation in vivo is dependent on ATR, and ATR phosphorylated FANCA on serine 1449 in vitro. Phosphorylation of FANCA on serine 1449 is a DNA damage-specific event that is downstream of ATR and is functionally important in the FA pathway.
Collins
Natalie B
NB
Department of Microbiology, University of Virginia School of Medicine, Charlottesville, USA.
Wilson
James B
JB
Bush
Thomas
T
Thomashevski
Andrei
A
Roberts
Kate J
KJ
Jones
Nigel J
NJ
Kupfer
Gary M
GM
D000477:Alkylating Agents / Q000633:toxicity; D064007:Ataxia Telangiectasia Mutated Proteins; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D005199:Fanconi Anemia / Q000235:genetics* / Q000378:metabolism*; D052217:Fanconi Anemia Complementation Group A Protein / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D016685:Mitomycin / Q000633:toxicity; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D012694:Serine / Q000378:metabolism
D000477:Alkylating Agents; D018797:Cell Cycle Proteins; C498471:FANCA protein, human; D052217:Fanconi Anemia Complementation Group A Protein; D012694:Serine; D016685:Mitomycin; C576597:ATR protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases