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SUMOylation regulates Rad18-mediated template switch

Nature volume 456, pages 915–920 (2008)Cite this article

Abstract

Replication by template switch is thought to mediate DNA damage-bypass and fillings of gaps. Gap-filling repair requires homologous recombination as well as Rad18- and Rad5-mediated proliferating cell nuclear antigen (PCNA) polyubiquitylation. However, it is unclear whether these processes are coordinated, and the physical evidence for Rad18–Rad5-dependent template switch at replication forks is still elusive. Here we show, using genetic and physical approaches, that in budding yeast (Saccharomyces cerevisiae) Rad18 is required for the formation of X-shaped sister chromatid junctions (SCJs) at damaged replication forks through a process involving PCNA polyubiquitylation and the ubiquitin-conjugating enzymes Mms2 and Ubc13. The Rad18–Mms2-mediated damage-bypass through SCJs requires the small ubiquitin-like modifier (SUMO)-conjugating enzyme Ubc9 and SUMOylated PCNA, and is coordinated with Rad51-dependent recombination events. We propose that the Rad18–Rad5–Mms2-dependent SCJs represent template switch events. Altogether, our results unmask a role for PCNA ubiquitylation and SUMOylation pathways in promoting transient damage-induced replication-coupled recombination events involving sister chromatids at replication forks.

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Figure 1: Rad18 promotes SCJ formation during replication of damaged templates, but its functionality requires SUMOylation.
Figure 2: Sgs1 functions to resolve the damage-induced SCJs formed by Rad18-Mms2-PCNA polyubiquitylation.
Figure 3: The contribution of PCNA SUMOylation in the formation of SCJs accumulating in sgs1 during replication of damaged templates.
Figure 4: Two pathways contributing to SCJ formation.

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Acknowledgements

We thank S. Jentsch and H. Ulrich for yeast strains, T. Enomoto for critical reading, and all members of our laboratories for helpful discussions. This work was supported by grants from the Associazione Italiana per la Ricerca sul Cancro to D.B., Association for International Cancer Research and European Community GENICA grant to M.F. and D.B., and partly by European Community DNA Repair grant, Telethon, MIUR, and Ministry of Health to M.F. D.B. was partly supported by the Buzzati-Traverso foundation.

Author Contributions D.B. conceived the project, designed and performed the experiments, and wrote the paper. F.V. quantified the 2D gels, performed data analysis and provided technical help. M.F. discussed the results, analysed the data, edited the manuscript and provided scientific advice and financial support.

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Authors and Affiliations

  1. IFOM, the FIRC Institute for Molecular Oncology Foundation, IFOM-IEO Campus, Via Adamello 16, 20139 Milan, Italy ,

    Dana Branzei, Fabio Vanoli & Marco Foiani

  2. Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, 20133 Milan, Italy

    Marco Foiani

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  1. Dana Branzei
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Correspondence to Dana Branzei.

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Branzei, D., Vanoli, F. & Foiani, M. SUMOylation regulates Rad18-mediated template switch. Nature 456, 915–920 (2008). https://doi.org/10.1038/nature07587

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