Abstract
The helicase-associated endonuclease for fork-structured DNA (Hef) is an archaeabacterial protein that processes blocked replication forks. Here we have isolated the vertebrate Hef ortholog and investigated its molecular function. Disruption of this gene in chicken DT40 cells results in genomic instability and sensitivity to DNA cross-links. The similarity of this phenotype to that of cells lacking the Fanconi anemia–related (FA) tumor-suppressor genes led us to investigate whether Hef functions in this pathway. Indeed, we found a genetic interaction between the FANCC and Hef genes. In addition, Hef is a component of the FA nuclear protein complex that facilitates its DNA damage–inducible chromatin localization and the monoubiquitination of the FA protein FANCD2. Notably, Hef interacts directly with DNA structures that are intermediates in DNA replication. This discovery sheds light on the origins, regulation and molecular function of the FA tumor-suppressor pathway in the maintenance of genome stability.
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Acknowledgements
G.M. was supported by a grant from Children's Leukaemia Trust, W.N. by a postdoctoral fellowship from Association of International Cancer Research, A.A. by a postdoctoral fellowship from Leukaemia Research Fund and F.L. by a grant from the Fanconi Anaemia Research Fund. We thank H. Joenje and J. DeWinter for the gift of the FANCA antibody and FA cell lines.
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Supplementary information
Supplementary Fig. 1
Evolutionary relationships in the Mph1-like helicases and XPF-like nucleases. (PDF 503 kb)
Supplementary Fig. 2
Clustal alignments of the vertebrate Hef orthologues. (PDF 1030 kb)
Supplementary Fig. 3
Disruption of the chicken HEF gene. (PDF 1022 kb)
Supplementary Fig. 4
Generation of an in-situ tagged FANCC and FANCF DT40 strain. (PDF 752 kb)
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Mosedale, G., Niedzwiedz, W., Alpi, A. et al. The vertebrate Hef ortholog is a component of the Fanconi anemia tumor-suppressor pathway. Nat Struct Mol Biol 12, 763–771 (2005). https://doi.org/10.1038/nsmb981
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DOI: https://doi.org/10.1038/nsmb981
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