Abstract
In budding yeast, DNA damage in G1 activates a Rad9-dependent checkpoint that targets the cyclin-dependent kinase (CDK) Cdc28 to delay G1 exit. After a transient arrest, cells may enter S phase before completing DNA repair. We used genetic analysis to identify the stress-responsive CDK Pho85, the cyclin Pho80 and the targeted transcription factors Pho4 and Swi5 as determinants of G1 checkpoint adaptation. Consistent with opposing roles for the Cdc28 inhibitor Sic1 in blocking G1 exit and Pho85 in targeting Sic1 for proteolysis, mutation of Sic1 curtails G1 checkpoint delay, whereas Pho85 inhibition after DNA damage promotes Sic1 stability. G1 checkpoint delay in mutants lacking both Sic1 and Pho4 is independent of Pho85 activity. These data establish a G1 checkpoint adaptation pathway where Pho85 mediates Pho4 downregulation and Sic1 degradation to release Cdc28 activity and promote onset of S phase.
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Acknowledgements
We dedicate this work in fond memory of our colleague, Cora Styles. We thank A. Hauser for technical assistance, P. Shah and D. Bishop for guidance with CHEF analysis and K. Shokat and E. O'Shea for generously sharing strains. This work was supported by grant R01 GM60443 from the US National Institutes of Health (S.J.K.) and grant PBZ-Min-015/P05/2004 from the Polish Ministry of Education and Science (R.W.). A.J. was supported by the University of Chicago Medical Scientist Training Program and an American Heart Association Greater Midwest predoctoral fellowship, K.K. by the University of Chicago's US National Institutes of Health Cancer Biology Training Grant and S.J.K. by a Leukemia & Lymphoma Society Scholar award.
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R.W. and A.J. conceived, performed and analyzed experiments and cowrote the manuscript; K.K. and F.S. performed and analyzed experiments; S.J.K. conceived and analyzed experiments and edited the manuscript.
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Wysocki, R., Javaheri, A., Kristjansdottir, K. et al. CDK Pho85 targets CDK inhibitor Sic1 to relieve yeast G1 checkpoint arrest after DNA damage. Nat Struct Mol Biol 13, 908–914 (2006). https://doi.org/10.1038/nsmb1139
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DOI: https://doi.org/10.1038/nsmb1139
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