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The DNA Pol ϵ stimulatory activity of Mrc1 is modulated by phosphorylation

Zhang, Zhong-Xin and Zhang, Jingjing and Cao, Qinhong and Campbell, Judith L. and Lou, Huiqiang (2018) The DNA Pol ϵ stimulatory activity of Mrc1 is modulated by phosphorylation. Cell Cycle, 17 (1). pp. 64-72. ISSN 1538-4101. PMCID PMC5815433. doi:10.1080/15384101.2017.1403680.

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DNA replication checkpoint (Mec1-Mrc1-Rad53 in budding yeast) is an evolutionarily conserved surveillance system to ensure proper DNA replication and genome stability in all eukaryotes. Compared to its well-known function as a mediator of replication checkpoint, the exact role of Mrc1 as a component of normal replication forks remains relatively unclear. In this study, we provide in vitro biochemical evidence to support that yeast Mrc1 is able to enhance the activity of DNA polymerase ϵ (Pol ϵ), the major leading strand replicase. Mrc1 can selectively bind avidly to primer/template DNA bearing a single-stranded region, but not to double-stranded DNA (dsDNA). Mutations of the lysine residues within basic patch 1 (BP1) compromise both DNA binding and polymerase stimulatory activities. Interestingly, Mrc1-3D, a mutant mimicking phosphorylation by the Hog1/MAPK kinase during the osmotic stress response, retains DNA binding but not polymerase stimulation. The stimulatory effect is also abrogated in Mrc1 purified from cells treated with hydroxyurea (HU), which elicits replication checkpoint activation. Taken together with previous findings, these results imply that under unperturbed condition, Mrc1 has a DNA synthesis stimulatory activity, which can be eliminated via Mrc1 phosphorylation in response to replication and/or osmotic stresses.

Item Type:Article
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URLURL TypeDescription CentralArticle
Lou, Huiqiang0000-0003-4465-6186
Additional Information:© 2017 Taylor & Francis. Received 18 Jul 2017, Accepted 29 Oct 2017, Accepted author version posted online: 20 Nov 2017, Published online: 21 Dec 2017. Funding: National Natural Science Foundation of China (NSFC) [grant number 31630005], [grant number 31770084], [grant number 31771382].
Funding AgencyGrant Number
National Natural Science Foundation of China31630005
National Natural Science Foundation of China31770084
National Natural Science Foundation of China31771382
Subject Keywords:Saccharomyces cerevisiae, DNA replication, Mrc1/Claspin, DNA polymerase ϵ, phosphorylation
Issue or Number:1
PubMed Central ID:PMC5815433
Record Number:CaltechAUTHORS:20171127-140130022
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Official Citation:Zhong-Xin Zhang, Jingjing Zhang, Qinhong Cao, Judith L. Campbell & Huiqiang Lou (2017) The DNA Pol ϵ stimulatory activity of Mrc1 is modulated by phosphorylation, Cell Cycle, 17:1, 64-72, DOI: 10.1080/15384101.2017.1403680
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:83449
Deposited By: Tony Diaz
Deposited On:27 Nov 2017 22:12
Last Modified:18 Mar 2022 22:14

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