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The vertebrate Hef ortholog is a component of the Fanconi anemia tumor-suppressor pathway

Nature Structural & Molecular Biology volume 12, pages 763771 (2005) | Download Citation

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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.

Author information

Author notes

    • Georgina Mosedale
    • , Wojciech Niedzwiedz
    •  & Arno Alpi

    These authors contributed equally to this work.

Affiliations

  1. Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.

    • Georgina Mosedale
    • , Wojciech Niedzwiedz
    • , Arno Alpi
    • , Franco Perrina
    • , Jose B Pereira-Leal
    • , Mark Johnson
    • , Frederic Langevin
    • , Paul Pace
    •  & Ketan J Patel
  2. Department of Medicine, Addenbrookes Hospital, Hills Road, Cambridge CB2 2QH, UK.

    • Ketan J Patel

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The authors declare no competing financial interests.

Corresponding author

Correspondence to Ketan J Patel.

Supplementary information

PDF files

  1. 1.

    Supplementary Fig. 1

    Evolutionary relationships in the Mph1-like helicases and XPF-like nucleases.

  2. 2.

    Supplementary Fig. 2

    Clustal alignments of the vertebrate Hef orthologues.

  3. 3.

    Supplementary Fig. 3

    Disruption of the chicken HEF gene.

  4. 4.

    Supplementary Fig. 4

    Generation of an in-situ tagged FANCC and FANCF DT40 strain.

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DOI

https://doi.org/10.1038/nsmb981

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