Halder, Swagata; Torrecilla, Ignacio; Burkhalter, Martin D.; Popović, Marta; Fielden, John; Vaz, Bruno; Oehler, Judith; Pilger, Domenic; Lessel, Davor; Wiseman, Katherine; Singh, Abhay Narayan; Vendrell, Iolanda; Fischer, Roman; Philipp, Melanie; Ramadan, Kristijan (2019) SPRTN protease and checkpoint kinase 1 cross-activation loop safeguards DNA replication. Nature Communications, 10 (1). pp. 1-18. ISSN 2041-1723
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Abstract
The SPRTN metalloprotease is essential for DNA-protein crosslink (DPC) repair and DNA replication in vertebrate cells. Cells deficient in SPRTN protease exhibit DPC-induced replication stress and genome instability, manifesting as premature ageing and liver cancer. Here, we provide a body of evidence suggesting that SPRTN activates the ATR-CHK1 phosphorylation signalling cascade during physiological DNA replication by proteolysis-dependent eviction of CHK1 from replicative chromatin. During this process, SPRTN proteolyses the Cterminal/inhibitory part of CHK1, liberating N-terminal CHK1 kinase active fragments. Simultaneously, CHK1 full length and its N-terminal fragments phosphorylate SPRTN at the C-terminal regulatory domain, which stimulates SPRTN recruitment to chromatin to promote unperturbed DNA replication fork progression and DPC repair. Our data suggest that a SPRTN-CHK1 cross-activation loop plays a part in DNA replication and protection from DNA replication stress. Finally, our results with purified components of this pathway further support the proposed model of a SPRTN-CHK1 cross-activation loop.
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| Additional Information: | The authors thank Cornelia Donow for technical assistance and excellent fish care. This work was supported by Medical Research Council-UK (MC_EX_MR/K022830/1) and Oxford Cancer Research Centre to K.R. S.H. was supported by Goodger and Schorstein Scholarship University of Oxford and a short-term postdoc fellowship by the European Institute of Innovation and Technology. The lab of M. Philipp is supported by Deutsche Forschungsgemeinschaft grant (PH144-4/-1) and the Boehringer Ingelheim Ulm University Biocenter, the lab of D. Lessel is supported by Deutsche Forschungsgemeinschaft grant (LE 4223/1) and the Deutsche Krebshilfe grant (70113348) and the lab of M. Popovic is supported by Croatian Science Foundation Installation Grant (UIP-2017-05-5258), Ruder Boskovic Institute (Zagreb, Croatia) and European Structural and Investment Funds STIM - REI project (KK.01.1.1.01.0003). Mass spectrometry analysis was performed in the TDI MS Laboratory led by Benedikt M. Kessler. The authors thank E. Despras and P.L. Kannouche for providing them with ssDNA staining protocols, O. Gileadi and J.A. Newman for their help with SPRTN protein purification and G. Dianov for Flag-CHK1 construct. The authors thank Anthony (Tony) Carr for critical reading of this manuscript and fruitful discussion. | ||||||||
| Uncontrolled Keywords: | SPRTN; Chk1; DNA replication; DNA-protein crosslinks | ||||||||
| Subjects: | NATURAL SCIENCES > Biology > Biochemistry and Molecular Biology BIOMEDICINE AND HEALTHCARE > Basic Medical Sciences |
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| Divisions: | Division for Marine and Enviromental Research | ||||||||
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| Depositing User: | Marta Popović | ||||||||
| Date Deposited: | 04 May 2020 06:51 | ||||||||
| URI: | http://fulir.irb.hr/id/eprint/5702 | ||||||||
| DOI: | 10.1038/s41467-019-11095-y |
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