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CKS Proteins Promote Checkpoint Recovery by Stimulating Phosphorylation of Treslin

Mu, Ruiling and Tat, John and Zamudio, Robert and Zhang, Yaoyang and Yates, John R., III and Kumagai, Akiko and Dunphy, William G. and Reed, Steven I. (2017) CKS Proteins Promote Checkpoint Recovery by Stimulating Phosphorylation of Treslin. Molecular and Cellular Biology, 37 (20). Art. No. e00344. ISSN 0270-7306. PMCID PMC5615187. https://resolver.caltech.edu/CaltechAUTHORS:20170731-094457327

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Abstract

CKS proteins are small (9-kDa) polypeptides that bind to a subset of the cyclin-dependent kinases. The two paralogs expressed in mammals, Cks1 and Cks2, share an overlapping function that is essential for early development. However, both proteins are frequently overexpressed in human malignancy. It has been shown that CKS protein overexpression overrides the replication stress checkpoint, promoting continued origin firing. This finding has led to the proposal that CKS protein-dependent checkpoint override allows premalignant cells to evade oncogene stress barriers, providing a causal link to oncogenesis. Here, we provide mechanistic insight into how overexpression of CKS proteins promotes override of the replication stress checkpoint. We show that CKS proteins greatly enhance the ability of Cdk2 to phosphorylate the key replication initiation protein treslin in vitro. Furthermore, stimulation of treslin phosphorylation does not occur by the canonical adapter mechanism demonstrated for other substrates, as cyclin-dependent kinase (CDK) binding-defective mutants are capable of stimulating treslin phosphorylation. This effect is recapitulated in vivo, where silencing of Cks1 and Cks2 decreases treslin phosphorylation, and overexpression of wild-type or CDK binding-defective Cks2 prevents checkpoint-dependent dephosphorylation of treslin. Finally, we provide evidence that the role of CKS protein-dependent checkpoint override involves recovery from checkpoint-mediated arrest of DNA replication.


Item Type:Article
Related URLs:
URLURL TypeDescription
https://doi.org/10.1128/MCB.00344-17DOIArticle
http://mcb.asm.org/content/37/20/e00344-17PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5615187PubMed CentralArticle
Additional Information:© 2017 American Society for Microbiology. Received 21 June 2017 Returned for modification 10 July 2017 Accepted 11 July 2017. Accepted manuscript posted online 24 July 2017. We thank Anindya Dutta for plasmids required for E. coli production of active cyclin E/Cdk2. This work was supported by NIH grants CA074224 and CA078343 to S.I.R., GM043974 and GM070891 to W.G.D., and P41 GM103533 to J.R.Y. We also thank the training grants NCATS 5 TL1 TR001114 and UL1 TR001114 and the Scripps Family Foundation and ARCS Foundation San Diego Chapter predoctoral fellowships for support of J.T. Author Contributions: S.I.R. supervised the research described. R.M., J.T., Y.Z., and S.I.R. designed and carried out experiments. R.Z. carried out experiments. A.K. and W.G.D. provided advice and reagents. S.I.R. wrote the manuscript, with editing help from A.K., W.G.D., J.T., and J.R.Y. J.R.Y. provided experimental resources.
Funders:
Funding AgencyGrant Number
NIHCA074224
NIHCA078343
NIHGM043974
NIHGM070891
NIHP41 GM103533
NIH5 TL1 TR001114
NIHUL1 TR001114
Scripps Family FoundationUNSPECIFIED
ARCS FoundationUNSPECIFIED
Subject Keywords:CKS protein; treslin; treslin phosphorylation; replication stress checkpoint; checkpoint recovery
Issue or Number:20
PubMed Central ID:PMC5615187
Record Number:CaltechAUTHORS:20170731-094457327
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170731-094457327
Official Citation:CKS Proteins Promote Checkpoint Recovery by Stimulating Phosphorylation of Treslin Ruiling Mu, John Tat, Robert Zamudio, Yaoyang Zhang, John R. Yates III, Akiko Kumagai, William G. Dunphy, and Steven I. Reed Mol. Cell. Biol. October 2017 37:20 15 e00344-17; Accepted manuscript posted online 24 July 2017, doi:10.1128/MCB.00344-17
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:79586
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Aug 2017 19:54
Last Modified:03 Oct 2019 18:21

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