About this item:
567 Views | 896 Downloads
Author Notes:
To whom correspondence should be addressed. Tel: +404 778 1758; Fax: +404 778 5520; Email: dsyu@emory.edu
The authors wish it to be known that, in their opinion, these authors should be considered as equal contributors.
We thank members of the D.S.Y. laboratory for helpful discussion.
Conflict of interest statement. None declared.
Subjects:
Research Funding:
Glenn Family Breast Cancer Foundation/Winship Cancer Institute [22868 to D.S.Y.]; National Institutes of Health [5P50CA128613 pilot award to D.S.Y.]; Georgia Research Alliance [11072 to D.S.Y.]. Funding for open access charge: Department Start-up Funds.
Keywords:
- Science & Technology
- Life Sciences & Biomedicine
- Biochemistry & Molecular Biology
- DNA-DAMAGE RESPONSE
- CELL-CYCLE ARREST
- CHECKPOINT CONTROL
- ASPERGILLUS-NIDULANS
- MITOTIC PROGRESSION
- ATR-ATRIP
- KINASE
- PROTEIN
- CHK1
- PHOSPHORYLATION
A gemcitabine sensitivity screen identifies a role for NEK9 in the replication stress response
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.
Copyright information:
© The Author(s) 2014.
This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits distribution, public display, and publicly performance, making multiple copies, distribution of derivative works, provided the original work is properly cited. This license requires credit be given to copyright holder and/or author, copyright and license notices be kept intact.
