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Mitotic phosphorylation of Bloom helicase at Thr182 is required for its proteasomal degradation and maintenance of chromosomal stability

Oncogene volume 35, pages 1025–1038 (2016)Cite this article

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Abstract

Mutations in Bloom helicase (BLM) lead to Bloom Syndrome (BS). BS is characterized by multiple clinical manifestations including predisposition to a wide spectrum of cancers. Studies have revealed the mechanism of BLM recruitment after stalled replication and its role during the repair of DNA damage. We now provide evidence that BLM undergoes K48-linked ubiquitylation and subsequent degradation during mitosis due to the E3 ligase, Fbw7α. Fbw7α carries out its function after GSK3β- and CDK2/cyclin A2-dependent phosphorylation events on Thr171 and Ser175 of BLM which lies within a well-defined phosphodegron, a sequence which is conserved in all primates. Phosphorylation on BLM Thr171 and Ser175 depends on prior phosphorylation at Thr182 by Chk1/Chk2. Thr182 phosphorylation not only controls BLM ubiquitylation and degradation during mitosis but is also a determinant for its localization on the ultrafine bridges. Consequently lack of Thr182 phosphorylation leads to multiple manifestations of chromosomal instability including increased levels of DNA damage, lagging chromatin, micronuclei formation, breaks and quadriradials. Hence Thr182 phosphorylation on BLM has two functions—it regulates BLM turnover during mitosis and also helps to maintain the chromosomal stability.

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Acknowledgements

We acknowledge Bruce Clurman, Markus Welcker, Jonathan Grim, Ian Hickson, Stephen Elledge, Thanos Halazonetis, Jerry Shay, Nathan Ellis, Bert Vogelstein for plasmids and cells, Simon Gemble for help with SCEs and UFB staining, Priyanka Modi for transfection experiments, Jyoti Kumari for lentivirus-mediated stable line generation and Richa Mudgal for bioinformatic analysis. SS acknowledges National Institute of Immunology core funds, Department of Biotechnology (DBT), India (BT/PR3148/AGR/36/706/2011); Science and Engineering Research Board, India (SR/SO/BB-0124/2013); Indo-French Centre for the Promotion of Advanced Research (IFCPAR) (IFC/4603-A/2011/1250) and Council of Scientific and Industrial Research (CSIR), India [37(1541)/12/EMR-II] for financial assistance.

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  1. S S Kharat, V Tripathi and A P Damodaran: These authors contributed equally to this work.

Authors and Affiliations

  1. National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi, India

    S S Kharat, V Tripathi, A P Damodaran, R Priyadarshini, S Chandra, S Tikoo, R Nandhakumar, V Srivastava, S Priya, M Hussain, S Kaur & S Sengupta

  2. 21st Century Biochemicals, 260 Cedar Hill Street, Marlboro, MA, USA

    J B Fishman

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Kharat, S., Tripathi, V., Damodaran, A. et al. Mitotic phosphorylation of Bloom helicase at Thr182 is required for its proteasomal degradation and maintenance of chromosomal stability. Oncogene 35, 1025–1038 (2016). https://doi.org/10.1038/onc.2015.157

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