Ethanol exposure increases mutation rate through error-prone polymerases

Karin Voordeckers; Camilla Colding; Lavinia Grasso; Benjamin Pardo; Lore Hoes; Jacek Kominek; Kim Gielens; Kaat Dekoster; Jonathan Gordon; Elisa Van der Zande; Peter Bircham; Toon Swings; Jan Michiels; Peter Van Loo; Sandra Nuyts; Philippe Pasero; Michael Lisby; Kevin J. Verstrepen

doi:10.1038/s41467-020-17447-3

Ethanol exposure increases mutation rate through error-prone polymerases

Karin Voordeckers, Camilla Colding, Lavinia Grasso, Benjamin Pardo, Lore Hoes, Jacek Kominek, Kim Gielens, Kaat Dekoster, Jonathan Gordon, Elisa Van der Zande, Peter Bircham, Toon Swings, Jan Michiels, Peter Van Loo, Sandra Nuyts, Philippe Pasero, Michael Lisby, Kevin J. Verstrepen

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Abstract

Ethanol is a ubiquitous environmental stressor that is toxic to all lifeforms. Here, we use the model eukaryote Saccharomyces cerevisiae to show that exposure to sublethal ethanol concentrations causes DNA replication stress and an increased mutation rate. Specifically, we find that ethanol slows down replication and affects localization of Mrc1, a conserved protein that helps stabilize the replisome. In addition, ethanol exposure also results in the recruitment of error-prone DNA polymerases to the replication fork. Interestingly, preventing this recruitment through mutagenesis of the PCNA/Pol30 polymerase clamp or deleting specific error-prone polymerases abolishes the mutagenic effect of ethanol. Taken together, this suggests that the mutagenic effect depends on a complex mechanism, where dysfunctional replication forks lead to recruitment of error-prone polymerases. Apart from providing a general mechanistic framework for the mutagenic effect of ethanol, our findings may also provide a route to better understand and prevent ethanol-associated carcinogenesis in higher eukaryotes.

Original language	English (US)
Article number	3664
Journal	Nature communications
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41467-020-17447-3
State	Published - Dec 1 2020

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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Voordeckers, K., Colding, C., Grasso, L., Pardo, B., Hoes, L., Kominek, J., Gielens, K., Dekoster, K., Gordon, J., Van der Zande, E., Bircham, P., Swings, T., Michiels, J., Van Loo, P., Nuyts, S., Pasero, P., Lisby, M., & Verstrepen, K. J. (2020). Ethanol exposure increases mutation rate through error-prone polymerases. Nature communications, 11(1), Article 3664. https://doi.org/10.1038/s41467-020-17447-3

Voordeckers, K, Colding, C, Grasso, L, Pardo, B, Hoes, L, Kominek, J, Gielens, K, Dekoster, K, Gordon, J, Van der Zande, E, Bircham, P, Swings, T, Michiels, J, Van Loo, P, Nuyts, S, Pasero, P, Lisby, M & Verstrepen, KJ 2020, 'Ethanol exposure increases mutation rate through error-prone polymerases', Nature communications, vol. 11, no. 1, 3664. https://doi.org/10.1038/s41467-020-17447-3

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title = "Ethanol exposure increases mutation rate through error-prone polymerases",

abstract = "Ethanol is a ubiquitous environmental stressor that is toxic to all lifeforms. Here, we use the model eukaryote Saccharomyces cerevisiae to show that exposure to sublethal ethanol concentrations causes DNA replication stress and an increased mutation rate. Specifically, we find that ethanol slows down replication and affects localization of Mrc1, a conserved protein that helps stabilize the replisome. In addition, ethanol exposure also results in the recruitment of error-prone DNA polymerases to the replication fork. Interestingly, preventing this recruitment through mutagenesis of the PCNA/Pol30 polymerase clamp or deleting specific error-prone polymerases abolishes the mutagenic effect of ethanol. Taken together, this suggests that the mutagenic effect depends on a complex mechanism, where dysfunctional replication forks lead to recruitment of error-prone polymerases. Apart from providing a general mechanistic framework for the mutagenic effect of ethanol, our findings may also provide a route to better understand and prevent ethanol-associated carcinogenesis in higher eukaryotes.",

author = "Karin Voordeckers and Camilla Colding and Lavinia Grasso and Benjamin Pardo and Lore Hoes and Jacek Kominek and Kim Gielens and Kaat Dekoster and Jonathan Gordon and {Van der Zande}, Elisa and Peter Bircham and Toon Swings and Jan Michiels and {Van Loo}, Peter and Sandra Nuyts and Philippe Pasero and Michael Lisby and Verstrepen, {Kevin J.}",

note = "Funding Information: We thank Patricia T.N. Van Dam (TU Delft) for technical assistance with the acetaldehyde HPLC measurements, as well as all CMPG members for discussions, Valmik Vyas for plasmid pV1382 and sharing the protocol for CRISPR, Judith Frydman for sharing the pESC-VHL-mCherry plasmid, and Boris Pfander for sharing plasmid pBP81. This work was financially supported by a grant from Research Foundation - Flanders (FWO)—grant number G090618N. K.V. acknowledges financial support from FWO by a postdoctoral fellowship (1249117 N). L.H. was supported by a PhD fellowship from FWO (11B6720N). J.K. was supported by a KU Leuven F + fellowship. M.L. is supported by The Independent Research Fund Denmark (FNU) and the Villum Foundation. Work in the lab of P.P. is funded by grants from the Agence Nationale pour la Recherche (ANR), the Institut National du Cancer(INCa), and the Ligue contre le Cancer ({\'e}quipe labellis{\'e}e). Research in the lab of K.J.V. is supported by KU Leuven Program Financing, European Research Council (ERC) Consolidator Grant CoG682009, Human Frontier Science (HFSP) program grant RGP0050/2013, Vlaams Instituut voor Biotechnologie (VIB), European Molecular Biology Organization (EMBO) Young Investigator program, FWO, and Agentschap voor Innovatie door Wetenschap en Technology (IWT). Research in the P.V.L. lab is supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001202), the UK Medical Research Council (FC001202), and the Wellcome Trust (FC001202). P.V.L. is a Winton Group Leader in recognition of the Winton Charitable Foundation{\textquoteright}s support towards the establishment of The Francis Crick Institute. Publisher Copyright: {\textcopyright} 2020, The Author(s).",

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doi = "10.1038/s41467-020-17447-3",

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T1 - Ethanol exposure increases mutation rate through error-prone polymerases

AU - Voordeckers, Karin

AU - Colding, Camilla

AU - Grasso, Lavinia

AU - Pardo, Benjamin

AU - Hoes, Lore

AU - Kominek, Jacek

AU - Gielens, Kim

AU - Dekoster, Kaat

AU - Gordon, Jonathan

AU - Van der Zande, Elisa

AU - Bircham, Peter

AU - Swings, Toon

AU - Michiels, Jan

AU - Van Loo, Peter

AU - Nuyts, Sandra

AU - Pasero, Philippe

AU - Lisby, Michael

AU - Verstrepen, Kevin J.

N1 - Funding Information: We thank Patricia T.N. Van Dam (TU Delft) for technical assistance with the acetaldehyde HPLC measurements, as well as all CMPG members for discussions, Valmik Vyas for plasmid pV1382 and sharing the protocol for CRISPR, Judith Frydman for sharing the pESC-VHL-mCherry plasmid, and Boris Pfander for sharing plasmid pBP81. This work was financially supported by a grant from Research Foundation - Flanders (FWO)—grant number G090618N. K.V. acknowledges financial support from FWO by a postdoctoral fellowship (1249117 N). L.H. was supported by a PhD fellowship from FWO (11B6720N). J.K. was supported by a KU Leuven F + fellowship. M.L. is supported by The Independent Research Fund Denmark (FNU) and the Villum Foundation. Work in the lab of P.P. is funded by grants from the Agence Nationale pour la Recherche (ANR), the Institut National du Cancer(INCa), and the Ligue contre le Cancer (équipe labellisée). Research in the lab of K.J.V. is supported by KU Leuven Program Financing, European Research Council (ERC) Consolidator Grant CoG682009, Human Frontier Science (HFSP) program grant RGP0050/2013, Vlaams Instituut voor Biotechnologie (VIB), European Molecular Biology Organization (EMBO) Young Investigator program, FWO, and Agentschap voor Innovatie door Wetenschap en Technology (IWT). Research in the P.V.L. lab is supported by the Francis Crick Institute, which receives its core funding from Cancer Research UK (FC001202), the UK Medical Research Council (FC001202), and the Wellcome Trust (FC001202). P.V.L. is a Winton Group Leader in recognition of the Winton Charitable Foundation’s support towards the establishment of The Francis Crick Institute. Publisher Copyright: © 2020, The Author(s).

PY - 2020/12/1

Y1 - 2020/12/1

N2 - Ethanol is a ubiquitous environmental stressor that is toxic to all lifeforms. Here, we use the model eukaryote Saccharomyces cerevisiae to show that exposure to sublethal ethanol concentrations causes DNA replication stress and an increased mutation rate. Specifically, we find that ethanol slows down replication and affects localization of Mrc1, a conserved protein that helps stabilize the replisome. In addition, ethanol exposure also results in the recruitment of error-prone DNA polymerases to the replication fork. Interestingly, preventing this recruitment through mutagenesis of the PCNA/Pol30 polymerase clamp or deleting specific error-prone polymerases abolishes the mutagenic effect of ethanol. Taken together, this suggests that the mutagenic effect depends on a complex mechanism, where dysfunctional replication forks lead to recruitment of error-prone polymerases. Apart from providing a general mechanistic framework for the mutagenic effect of ethanol, our findings may also provide a route to better understand and prevent ethanol-associated carcinogenesis in higher eukaryotes.

AB - Ethanol is a ubiquitous environmental stressor that is toxic to all lifeforms. Here, we use the model eukaryote Saccharomyces cerevisiae to show that exposure to sublethal ethanol concentrations causes DNA replication stress and an increased mutation rate. Specifically, we find that ethanol slows down replication and affects localization of Mrc1, a conserved protein that helps stabilize the replisome. In addition, ethanol exposure also results in the recruitment of error-prone DNA polymerases to the replication fork. Interestingly, preventing this recruitment through mutagenesis of the PCNA/Pol30 polymerase clamp or deleting specific error-prone polymerases abolishes the mutagenic effect of ethanol. Taken together, this suggests that the mutagenic effect depends on a complex mechanism, where dysfunctional replication forks lead to recruitment of error-prone polymerases. Apart from providing a general mechanistic framework for the mutagenic effect of ethanol, our findings may also provide a route to better understand and prevent ethanol-associated carcinogenesis in higher eukaryotes.

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JO - Nature communications

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Ethanol exposure increases mutation rate through error-prone polymerases

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