TY - JOUR
T1 - Fanconi anemia proteins are required to prevent accumulation of replication-associated DNA double-strand breaks
AU - Sobeck, Alexandra
AU - Stone, Stacie
AU - Costanzo, Vincenzo
AU - De Graaf, Bendert
AU - Reuter, Tanja
AU - De Winter, Johan
AU - Wallisch, Michael
AU - Akkari, Yassmine
AU - Olson, Susan
AU - Wang, Weidong
AU - Joenje, Hans
AU - Christian, Jan L.
AU - Lupardus, Patrick J.
AU - Cimprich, Karlene A.
AU - Gautier, Jean
AU - Hoatlin, Maureen E.
PY - 2006/1
Y1 - 2006/1
N2 - Fanconi anemia (FA) is a multigene cancer susceptibility disorder characterized by cellular hypersensitivity to DNA interstrand cross-linking agents such as mitomycin C (MMC). FA proteins are suspected to function at the interface between cell cycle checkpoints, DNA repair, and DNA replication. Using replicating extracts from Xenopus eggs, we developed cell-free assays for FA proteins (xFA). Recruitment of the xFA core complex and xFANCD2 to chromatin is strictly dependent on replication initiation, even in the presence of MMC indicating specific recruitment to DNA lesions encountered by the replication machinery. The increase in xFA chromatin binding following treatment with MMC is part of a caffeine-sensitive S-phase checkpoint that is controlled by xATR. Recruitment of xFANCD2, but not xFANCA, is dependent on the xATR-xATR- interacting protein (xATRIP) complex. Immunodepletion of either xFANCA or xFANCD2 from egg extracts results in accumulation of chromosomal DNA breaks during replicative synthesis. Our results suggest coordinated chromatin recruitment of xFA proteins in response to replication-associated DNA lesions and indicate that xFA proteins function to prevent the accumulation of DNA breaks that arise during unperturbed replication.
AB - Fanconi anemia (FA) is a multigene cancer susceptibility disorder characterized by cellular hypersensitivity to DNA interstrand cross-linking agents such as mitomycin C (MMC). FA proteins are suspected to function at the interface between cell cycle checkpoints, DNA repair, and DNA replication. Using replicating extracts from Xenopus eggs, we developed cell-free assays for FA proteins (xFA). Recruitment of the xFA core complex and xFANCD2 to chromatin is strictly dependent on replication initiation, even in the presence of MMC indicating specific recruitment to DNA lesions encountered by the replication machinery. The increase in xFA chromatin binding following treatment with MMC is part of a caffeine-sensitive S-phase checkpoint that is controlled by xATR. Recruitment of xFANCD2, but not xFANCA, is dependent on the xATR-xATR- interacting protein (xATRIP) complex. Immunodepletion of either xFANCA or xFANCD2 from egg extracts results in accumulation of chromosomal DNA breaks during replicative synthesis. Our results suggest coordinated chromatin recruitment of xFA proteins in response to replication-associated DNA lesions and indicate that xFA proteins function to prevent the accumulation of DNA breaks that arise during unperturbed replication.
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U2 - 10.1128/MCB.26.2.425-437.2006
DO - 10.1128/MCB.26.2.425-437.2006
M3 - Article
C2 - 16382135
AN - SCOPUS:30644459206
SN - 0270-7306
VL - 26
SP - 425
EP - 437
JO - Molecular and cellular biology
JF - Molecular and cellular biology
IS - 2
ER -