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Sgs1 helicase is required for efficient PCNA monoubiquitination and translesion DNA synthesis in Saccharomyces cerevisiae

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Abstract

DNA-damage tolerance (DDT) is employed by eukaryotes to deal with replication blocks on the template strand, and is divided into two parallel pathways that are activated by sequential ubiquitination of proliferating cell nuclear antigen (PCNA) at the Lys164 residue. Rad6–Rad18-mediated PCNA monoubiquitination promotes translesion DNA synthesis (TLS) and the monoubiquitinated PCNA can be further polyubiquitinated by an Mms2-Ubc13-Rad5 complex, leading to error-free lesion bypass. We previously reported that the DNA helicase Sgs1 is required for error-free lesion bypass, probably through the double-Holliday junction migration and subsequent resolution. Surprisingly, a synthetic genetic array (SGA) screen using rev1 and rev3 as baits did not reveal an anticipated synthetic effect with sgs1, indicating a possible involvement of Sgs1 in TLS. Here, we report detailed genetic analyses demonstrating that Sgs1 plays a key role in efficient TLS and that it is probably required for the signaling of DNA damage leading to PCNA monoubiquitination. These studies collectively illustrate that Sgs1 participates in both branches of DDT and possibly plays a role in pathway choice.

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Acknowledgements

We wish to thank Dr. I. Hickson for SGS1-related reagents and other Xiao laboratory members for technical assistance and helpful discussion. This work was supported by the National Natural Science Foundation of China operating grant 31670068, Chinese National 973 Project 2013CB911003 and Natural Sciences and Engineering Research Council of Canada Discovery Grants RGPIN-2014-04580 to W.X.

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Authors and Affiliations

  1. Beijing Key Laboratory of DNA Damage Responses and College of Life Sciences, Capital Normal University, Beijing, 100048, China

    Fangfang Li, Li Fan & Wei Xiao

  2. Department of Microbiology and Immunology, University of Saskatchewan, Saskatoon, SK, S7N 5E5, Canada

    Lindsay G. Ball, Michelle Hanna & Wei Xiao

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  1. Fangfang Li
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  2. Lindsay G. Ball
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  3. Li Fan
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  5. Wei Xiao
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Correspondence to Wei Xiao.

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Communicated by M. Kupiec.

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Li, F., Ball, L.G., Fan, L. et al. Sgs1 helicase is required for efficient PCNA monoubiquitination and translesion DNA synthesis in Saccharomyces cerevisiae . Curr Genet 64, 459–468 (2018). https://doi.org/10.1007/s00294-017-0753-0

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