Abstract
The DNA replication machinery invariably encounters obstacles that slow replication fork progression, and threaten to prevent complete replication and faithful segregation of sister chromatids. The resulting replication stress activates ATR, the major kinase involved in resolving impaired DNA replication. In addition, replication stress also activates the related kinase ATM, which is required to prevent mitotic segregation errors. However, the molecular mechanism of ATM activation by replication stress is not defined. Here, we show that monoubiquitinated Proliferating Cell Nuclear Antigen (PCNA), a marker of stalled replication forks, interacts with the ATM cofactor ATMIN via WRN-interacting protein 1 (WRNIP1). ATMIN, WRNIP1 and RAD18, the E3 ligase responsible for PCNA monoubiquitination, are specifically required for ATM signalling and 53BP1 focus formation induced by replication stress, not ionising radiation. Thus, WRNIP1 connects PCNA monoubiquitination with ATMIN/ATM to activate ATM signalling in response to replication stress and contribute to the maintenance of genomic stability.
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Change history
28 July 2016
This article has been corrected since Advance Online Publication and a corrigendum is also printed in this issue
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Acknowledgements
Support was given from Mass Spectrometry, Bioinformatics and Biostatistics and the Equipment Park in the Cancer Research UK London Research Institute (now part of the Francis Crick Institute). We are grateful to JL Parker in the Ulrich laboratory for preparing ubiquitinated PCNA. R Bish, I Dikic, M Myers, E Soutoglou and A Lehmann kindly provided experimental materials. NK is funded by the Breast Cancer Research Foundation. TZ was financially supported by an A*STAR NSS-PhD scholarship (Singapore). This work was supported by an ERC grant (281661 ATMINDDR) to AB.
Author contributions
NK designed and performed most of the experiments and analysed the data. TZ, RAB, AC, JC and CDC performed experiments. EG, HNP, EA and CS provided technical assistance. SS, LG and HDU provided reagents and technical assistance. NK and AB wrote the manuscript.
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Kanu, N., Zhang, T., Burrell, R. et al. RAD18, WRNIP1 and ATMIN promote ATM signalling in response to replication stress. Oncogene 35, 4009–4019 (2016). https://doi.org/10.1038/onc.2015.427
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DOI: https://doi.org/10.1038/onc.2015.427
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