Kinesin Kif2C in regulation of DNA double strand break dynamics and repair.

Songli Zhu,Aimin Peng,Feifei Wang,Benjamin H Kwok,Ling Wang,Tadayoshi Bessho,Mohammadjavad Paydar,Benoit Barrette,Yanqiu Li

doi:10.7554/elife.53402

Abstract

DNA double strand breaks (DSBs) have detrimental effects on cell survival and genomic stability, and are related to cancer and other human diseases. In this study, we identified microtubule-depolymerizing kinesin Kif2C as a protein associated with DSB-mimicking DNA templates and known DSB repair proteins in Xenopus egg extracts and mammalian cells. The recruitment of Kif2C to DNA damage sites was dependent on both PARP and ATM activities. Kif2C knockdown or knockout led to accumulation of endogenous DNA damage, DNA damage hypersensitivity, and reduced DSB repair via both NHEJ and HR. Interestingly, Kif2C depletion, or inhibition of its microtubule depolymerase activity, reduced the mobility of DSBs, impaired the formation of DNA damage foci, and decreased the occurrence of foci fusion and resolution. Taken together, our study established Kif2C as a new player of the DNA damage response, and presented a new mechanism that governs DSB dynamics and repair.

Highlights

DNA damage is frequently induced by both endogenous metabolic products and exogenous genotoxic agents
As described in our previous study (Zhu et al, 2017), we utilized DNA double strand breaks (DSBs)-mimicking dA-dT oligonucleotides to isolate potential DNA damage-associated proteins in Xenopus egg extract, a cell-free system well-defined for studying DNA damage repair and signaling (Guo et al, 1999; Lupardus et al, 2007)
Along with Ku70, PARP1, RPA, and many other factors known to be involved in DSB repair, Kif2C was proteomically identified as a co-precipitated protein of dA-dT

Summary

Introduction

DNA damage is frequently induced by both endogenous metabolic products and exogenous genotoxic agents. NHEJ directly re-ligates the two broken ends of a DSB, and is accessible throughout the entire interphase (Davis and Chen, 2013; Lieber, 2010) In addition to these core pathways of DSB repair, the spatiotemporal regulation of DSBs has emerged as a new aspect of DNA repair (Amitai et al, 2017; Chuang et al, 2006; Chung et al, 2015; Hauer and Gasser, 2017; Krawczyk et al, 2012; Lemaıtre and Soutoglou, 2015; Levi et al, 2005; Lottersberger et al, 2015; Marcomini et al, 2018; Marnef and Legube, 2017; Mine-Hattab and Rothstein, 2013; Neumaier et al, 2012; Schrank et al, 2018).

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