A gemcitabine sensitivity screen identifies a role for NEK9 in the replication stress response

Scott C. Smith, Aleksandra V. Petrova, Matthew Z. Madden, Hongyan Wang, Yunfeng Pan, Matthew D. Warren, Claire W. Hardy, Dong Liang, Elaine A. Liu, M. Hope Robinson, Soumon Rudra, Jie Wang, Shahrzad Ehdaivand, Mylin A. Torres, Ya Wang, David S. Yu

Research output: Contribution to journalArticle

19 Scopus citations

Abstract

The Replication Stress Response (RSR) is a signaling network that recognizes challenges to DNA replication and coordinates diverse DNA repair and cell-cycle checkpoint pathways. Gemcitabine is a nucleoside analogue that causes cytotoxicity by inducing DNA replication blocks. Using a synthetic lethal screen of a RNAi library of nuclear enzymes to identify genes that when silenced cause gemcitabine sensitization or resistance in human triple-negative breast cancer cells, we identified NIMA (never in mitosis gene A)-related kinase 9 (NEK9) as a key component of the RSR. NEK9 depletion in cells leads to replication stress hypersensitivity, spontaneous accumulation of DNA damage and RPA70 foci, and an impairment in recovery from replication arrest. NEK9 protein levels also increase in response to replication stress. NEK9 complexes with CHK1, and moreover, NEK9 depletion impairs CHK1 autophosphorylation and kinase activity in response to replication stress. Thus, NEK9 is a critical component of the RSR that promotes CHK1 activity, maintaining genome integrity following challenges to DNA replication.

Original languageEnglish (US)
Pages (from-to)11517-11527
Number of pages11
JournalNucleic acids research
Volume42
Issue number18
DOIs
Publication statusPublished - Oct 13 2014

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ASJC Scopus subject areas

  • Genetics

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Smith, S. C., Petrova, A. V., Madden, M. Z., Wang, H., Pan, Y., Warren, M. D., ... Yu, D. S. (2014). A gemcitabine sensitivity screen identifies a role for NEK9 in the replication stress response. Nucleic acids research, 42(18), 11517-11527. https://doi.org/10.1093/nar/gku840