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RASSF1A–LATS1 signalling stabilizes replication forks by restricting CDK2-mediated phosphorylation of BRCA2

Nature Cell Biology volume 16, pages 962971 (2014) | Download Citation

  • An Erratum to this article was published on 27 March 2015

This article has been updated

Abstract

Genomic instability is a key hallmark of cancer leading to tumour heterogeneity and therapeutic resistance. BRCA2 has a fundamental role in error-free DNA repair but also sustains genome integrity by promoting RAD51 nucleofilament formation at stalled replication forks. CDK2 phosphorylates BRCA2 (pS3291-BRCA2) to limit stabilizing contacts with polymerized RAD51; however, how replication stress modulates CDK2 activity and whether loss of pS3291-BRCA2 regulation results in genomic instability of tumours are not known. Here we demonstrate that the Hippo pathway kinase LATS1 interacts with CDK2 in response to genotoxic stress to constrain pS3291-BRCA2 and support RAD51 nucleofilaments, thereby maintaining genomic fidelity during replication stalling. We also show that LATS1 forms part of an ATR-mediated response to replication stress that requires the tumour suppressor RASSF1A. Importantly, perturbation of the ATR–RASSF1A–LATS1 signalling axis leads to genomic defects associated with loss of BRCA2 function and contributes to genomic instability and ‘BRCA-ness’ in lung cancers.

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Change history

  • 11 March 2015

    In the original version of this Article, the bar chart in Fig. 6 was incorrect. This has now been corrected in all online versions of this Article.

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Acknowledgements

The authors would like to acknowledge T. Helleday for helpful discussions, M. Woodcock for fluorescence-activated cell sorting analysis and A. G. Abraham for useful comments on the manuscript. Lats1−/− MEFs were provided by T. Xu and myc–LATS1 constructs by H. Silljé. This work was funded by Cancer Research UK A19277, Vertex and the Medical Research Council (MRC).

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Affiliations

  1. CRUK/MRC Oxford Institute, Department of Oncology, University of Oxford, Oxford OX3 7DQ, UK

    • Dafni-Eleftheria Pefani
    • , Robert Latusek
    • , Isabel Pires
    • , Anna M. Grawenda
    • , Karen S. Yee
    • , Garth Hamilton
    • , Ester M. Hammond
    •  & Eric O’Neill
  2. The Wellcome Trust Sanger Institute, Hinxton, Cambridge CB10 1HH, UK

    • Louise van der Weyden
  3. Dunn School of Pathology, South Parks Road, University of Oxford, Oxford OX1 3RE, UK

    • Fumiko Esashi

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Contributions

D-E.P. designed and performed most of the experiments, analysed data and contributed to writing the paper; R.L. performed initial experiments for LATS1–CDK2 interaction, γH2AX analysis, pBRCA2 western blots and I-SceI assay; I.P. transferred the DNA fibre technology; K.S.Y. performed initial immunoprecipitation experiments for RASSF1A effect on LATS1–CDK2 interaction. G.H. performed the in vitro kinase assays; A.M.G. performed the TCGA analysis; L.v.d.W. isolated Rassf1A−/− mouse embryonic fibroblasts; F.E. provided BRCA2 reagents and advice; E.M.H. helped with reagents and advice; E.O’N. conceived, coordinated and supervised the project, designed experiments, analysed data and wrote the paper.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Eric O’Neill.

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https://doi.org/10.1038/ncb3035

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