Abstract
Fanconi anemia (FA) is characterized by cellular hypersensitivity to DNA crosslinking agents, but how the Fanconi pathway protects cells from DNA crosslinks and whether FA proteins act directly on crosslinks remain unclear. We developed a chromatin-IP-based strategy termed eChIP and detected association of multiple FA proteins with DNA crosslinks in vivo. Interdependence analyses revealed that crosslink-specific enrichment of various FA proteins is controlled by distinct mechanisms. BRCA-related FA proteins (BRCA2, FANCJ/BACH1, and FANCN/PALB2), but not FA core and I/D2 complexes, require replication for their crosslink association. FANCD2, but not FANCJ and FANCN, requires the FA core complex for its recruitment. FA core complex requires nucleotide excision repair proteins XPA and XPC for its association. Consistent with the distinct recruitment mechanism, recombination-independent crosslink repair was inversely affected in cells deficient of FANC-core versus BRCA-related FA proteins. Thus, FA proteins participate in distinct DNA damage response mechanisms governed by DNA replication status.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, N.I.H., Intramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Apoptosis Regulatory Proteins
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BRCA2 Protein / genetics
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BRCA2 Protein / metabolism*
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Basic-Leucine Zipper Transcription Factors / genetics
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Basic-Leucine Zipper Transcription Factors / metabolism*
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Breast Neoplasms / genetics*
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Breast Neoplasms / metabolism
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Breast Neoplasms / pathology
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Cell Line, Tumor
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Chromatin Immunoprecipitation / methods
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Cross-Linking Reagents / pharmacology
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DNA Damage*
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DNA Replication*
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DNA-Binding Proteins / metabolism
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Fanconi Anemia Complementation Group D2 Protein / genetics
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Fanconi Anemia Complementation Group D2 Protein / metabolism*
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Fanconi Anemia Complementation Group N Protein
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Fanconi Anemia Complementation Group Proteins / genetics
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Fanconi Anemia Complementation Group Proteins / metabolism*
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Female
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Ficusin / pharmacology
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Gene Expression Regulation, Neoplastic
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Humans
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Mutation
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism*
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Plasmids / metabolism
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Recombination, Genetic
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Time Factors
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Tumor Suppressor Proteins / genetics
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Tumor Suppressor Proteins / metabolism*
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Xeroderma Pigmentosum Group A Protein / metabolism
Substances
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Apoptosis Regulatory Proteins
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BACH1 protein, human
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BLID protein, human
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BRCA2 Protein
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BRCA2 protein, human
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Basic-Leucine Zipper Transcription Factors
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Cross-Linking Reagents
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DNA-Binding Proteins
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FANCD2 protein, human
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Fanconi Anemia Complementation Group D2 Protein
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Fanconi Anemia Complementation Group N Protein
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Fanconi Anemia Complementation Group Proteins
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Nuclear Proteins
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PALB2 protein, human
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Tumor Suppressor Proteins
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XPA protein, human
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Xeroderma Pigmentosum Group A Protein
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XPC protein, human
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Ficusin