Fanconi anemia proteins stabilize replication forks.
Fanconi anemia (FA) is a recessive genetic disorder characterized by hypersensitivity to crosslinking agents that has been attributed to defects in DNA repair and/or replication. FANCD2 and the FA core complex bind to chromatin during DNA replication; however, the role of FA proteins during replication is unknown. Using Xenopus cell-free extracts, we show that FANCL depletion results in defective DNA replication restart following treatment with camptothecin, a drug that results in DSBs during DNA replication. This defect is more pronounced following treatment with mitomycin C, presumably because of an additional role of the FA pathway in DNA crosslink repair. Moreover, we show that chromatin binding of FA core complex proteins during DNA replication follows origin assembly and origin firing and is dependent on the binding of RPA to ssDNA while FANCD2 additionally requires ATR, consistent with FA proteins acting at replication forks. Together, our data suggest that FA proteins play a role in replication restart at collapsed replication forks.
Wang
Lily Chien
LC
Institute for Cancer Genetics, Columbia University, New York, NY 10032, USA.
Stone
Stacie
S
Hoatlin
Maureen Elizabeth
ME
Gautier
Jean
J
D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D016590:Aphidicolin / Q000494:pharmacology; D064007:Ataxia Telangiectasia Mutated Proteins; D002166:Camptothecin / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D002474:Cell-Free System; D002843:Chromatin / Q000378:metabolism*; D004249:DNA Damage; D004260:DNA Repair; D004261:DNA Replication; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005199:Fanconi Anemia; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000502:physiology*; D052240:Fanconi Anemia Complementation Group L Protein / Q000502:physiology*; D016685:Mitomycin / Q000494:pharmacology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D029867:Xenopus Proteins / Q000378:metabolism; D014982:Xenopus laevis
D000903:Antibiotics, Antineoplastic; D000972:Antineoplastic Agents, Phytogenic; D018797:Cell Cycle Proteins; D002843:Chromatin; D004791:Enzyme Inhibitors; D052236:Fanconi Anemia Complementation Group D2 Protein; D029867:Xenopus Proteins; D016590:Aphidicolin; D016685:Mitomycin; D052240:Fanconi Anemia Complementation Group L Protein; C576898:Atr protein, Xenopus; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases; D002166:Camptothecin
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