Fanconi anemia proteins are required to prevent accumulation of replication-associated DNA double-strand breaks

Alexandra Sobeck, Stacie Stone, Vincenzo Costanzo, Bendert De Graaf, Tanja Reuter, Johan De Winter, Michael Wallisch, Yassmine Akkari, Susan Olson, Weidong Wang, Hans Joenje, Jan L. Christian, Patrick J. Lupardus, Karlene A. Cimprich, Jean Gautier, Maureen E. Hoatlin

Research output: Contribution to journalArticle

82 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)425-437
Number of pages13
JournalMolecular and cellular biology
Volume26
Issue number2
DOIs
StatePublished - Jan 1 2006

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Fanconi anemia proteins are required to prevent accumulation of replication-associated DNA double-strand breaks'. Together they form a unique fingerprint.

  • Cite this

    Sobeck, A., Stone, S., Costanzo, V., De Graaf, B., Reuter, T., De Winter, J., Wallisch, M., Akkari, Y., Olson, S., Wang, W., Joenje, H., Christian, J. L., Lupardus, P. J., Cimprich, K. A., Gautier, J., & Hoatlin, M. E. (2006). Fanconi anemia proteins are required to prevent accumulation of replication-associated DNA double-strand breaks. Molecular and cellular biology, 26(2), 425-437. https://doi.org/10.1128/MCB.26.2.425-437.2006