Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex

Lovely Jael Paul Solomon Devakumar; Christl Gaubitz; Victoria Lundblad; Brian A. Kelch; Takashi Kubota

doi:10.1093/nar/gkz441

Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex

Lovely Jael Paul Solomon Devakumar, Christl Gaubitz, Victoria Lundblad, Brian A. Kelch, Takashi Kubota^* (Corresponding Author)

^*Corresponding author for this work

Medical Sciences

Research output: Contribution to journal › Article › peer-review

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Abstract

Proliferating cell nuclear antigen (PCNA) is a sliding clamp that acts as a central co-ordinator for mismatch repair (MMR) as well as DNA replication. Loss of Elg1, the major subunit of the PCNA unloader complex, causes over-accumulation of PCNA on DNA and also increases mutation rate, but it has been unclear if the two effects are linked. Here we show that timely removal of PCNA from DNA by the Elg1 complex is important to prevent mutations. Although premature unloading of PCNA generally increases mutation rate, the mutator phenotype of elg1Δ is attenuated by PCNA mutants PCNA-R14E and PCNA-D150E that spontaneously fall off DNA. In contrast, the elg1Δ mutator phenotype is exacerbated by PCNA mutants that accumulate on DNA due to enhanced electrostatic PCNA–DNA interactions. Epistasis analysis suggests that PCNA over-accumulation on DNA interferes with both MMR and MMR-independent process(es). In elg1Δ, over-retained PCNA hyper-recruits the Msh2–Msh6 mismatch recognition complex through its PCNA-interacting peptide motif, causing accumulation of MMR intermediates. Our results suggest that PCNA retention controlled by the Elg1 complex is critical for efficient MMR: PCNA needs to be on DNA long enough to enable MMR, but if it is retained too long it interferes with downstream repair steps.

Original language	English
Pages (from-to)	6826-6841
Number of pages	16
Journal	Nucleic Acids Research
Volume	47
Issue number	13
Early online date	22 May 2019
DOIs	https://doi.org/10.1093/nar/gkz441
Publication status	Published - 26 Jul 2019

Bibliographical note

ACKNOWLEDGEMENTS
We thank Richard Kolodner and Eric Alani for strains and plasmids. We thank Anne Donaldson, Alexander Lorenz and Catherine Johnson from University of Aberdeen for careful reading of the manuscript. We thank Annabelle Duff and Veronika Petrova for assisting with the mutation rate assays, and Duru Cosar for assisting with crystal structure analysis. We appreciate assistance from staff of the Microscopy and Histology Core Facility and the qPCR facility at the University of Aberdeen.

FUNDING
Medical Research Council (MRC) Career Development Fellowship [L019698/1 to T.K.]; American Cancer Society Research Scholar Award [Grant #440685 to B.A.K.]; National Institute of General Medical Sciences [R01 GM127776 to B.A.K.]; National Institutes of Health grant [R01 GM106060 to V.L.]. Swiss National Science Foundation Postdoc Mobility Fellowship (to C.G.). Funding for open access charge: Medical Research Council via University of Aberdeen Open Access Fund.

Keywords

REPLICATION FACTOR-C
CELL NUCLEAR ANTIGEN
SACCHAROMYCES-CEREVISIAE
GENOME STABILITY
S-PHASE
TRANSLESION SYNTHESIS
TELOMERE LENGTH
PROTEIN
FORMS
INSTABILITY

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10.1093/nar/gkz441Licence: CC BY

Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex
© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
Final published version, 5.96 MBLicence: CC BY

Cite this

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title = "Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex",

abstract = "Proliferating cell nuclear antigen (PCNA) is a sliding clamp that acts as a central co-ordinator for mismatch repair (MMR) as well as DNA replication. Loss of Elg1, the major subunit of the PCNA unloader complex, causes over-accumulation of PCNA on DNA and also increases mutation rate, but it has been unclear if the two effects are linked. Here we show that timely removal of PCNA from DNA by the Elg1 complex is important to prevent mutations. Although premature unloading of PCNA generally increases mutation rate, the mutator phenotype of elg1Δ is attenuated by PCNA mutants PCNA-R14E and PCNA-D150E that spontaneously fall off DNA. In contrast, the elg1Δ mutator phenotype is exacerbated by PCNA mutants that accumulate on DNA due to enhanced electrostatic PCNA–DNA interactions. Epistasis analysis suggests that PCNA over-accumulation on DNA interferes with both MMR and MMR-independent process(es). In elg1Δ, over-retained PCNA hyper-recruits the Msh2–Msh6 mismatch recognition complex through its PCNA-interacting peptide motif, causing accumulation of MMR intermediates. Our results suggest that PCNA retention controlled by the Elg1 complex is critical for efficient MMR: PCNA needs to be on DNA long enough to enable MMR, but if it is retained too long it interferes with downstream repair steps.",

keywords = "REPLICATION FACTOR-C, CELL NUCLEAR ANTIGEN, SACCHAROMYCES-CEREVISIAE, GENOME STABILITY, S-PHASE, TRANSLESION SYNTHESIS, TELOMERE LENGTH, PROTEIN, FORMS, INSTABILITY",

author = "{Paul Solomon Devakumar}, {Lovely Jael} and Christl Gaubitz and Victoria Lundblad and Kelch, {Brian A.} and Takashi Kubota",

note = "ACKNOWLEDGEMENTS We thank Richard Kolodner and Eric Alani for strains and plasmids. We thank Anne Donaldson, Alexander Lorenz and Catherine Johnson from University of Aberdeen for careful reading of the manuscript. We thank Annabelle Duff and Veronika Petrova for assisting with the mutation rate assays, and Duru Cosar for assisting with crystal structure analysis. We appreciate assistance from staff of the Microscopy and Histology Core Facility and the qPCR facility at the University of Aberdeen. FUNDING Medical Research Council (MRC) Career Development Fellowship [L019698/1 to T.K.]; American Cancer Society Research Scholar Award [Grant #440685 to B.A.K.]; National Institute of General Medical Sciences [R01 GM127776 to B.A.K.]; National Institutes of Health grant [R01 GM106060 to V.L.]. Swiss National Science Foundation Postdoc Mobility Fellowship (to C.G.). Funding for open access charge: Medical Research Council via University of Aberdeen Open Access Fund.",

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doi = "10.1093/nar/gkz441",

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pages = "6826--6841",

journal = "Nucleic Acids Research",

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T1 - Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex

AU - Paul Solomon Devakumar, Lovely Jael

AU - Gaubitz, Christl

AU - Lundblad, Victoria

AU - Kelch, Brian A.

AU - Kubota, Takashi

N1 - ACKNOWLEDGEMENTS We thank Richard Kolodner and Eric Alani for strains and plasmids. We thank Anne Donaldson, Alexander Lorenz and Catherine Johnson from University of Aberdeen for careful reading of the manuscript. We thank Annabelle Duff and Veronika Petrova for assisting with the mutation rate assays, and Duru Cosar for assisting with crystal structure analysis. We appreciate assistance from staff of the Microscopy and Histology Core Facility and the qPCR facility at the University of Aberdeen. FUNDING Medical Research Council (MRC) Career Development Fellowship [L019698/1 to T.K.]; American Cancer Society Research Scholar Award [Grant #440685 to B.A.K.]; National Institute of General Medical Sciences [R01 GM127776 to B.A.K.]; National Institutes of Health grant [R01 GM106060 to V.L.]. Swiss National Science Foundation Postdoc Mobility Fellowship (to C.G.). Funding for open access charge: Medical Research Council via University of Aberdeen Open Access Fund.

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N2 - Proliferating cell nuclear antigen (PCNA) is a sliding clamp that acts as a central co-ordinator for mismatch repair (MMR) as well as DNA replication. Loss of Elg1, the major subunit of the PCNA unloader complex, causes over-accumulation of PCNA on DNA and also increases mutation rate, but it has been unclear if the two effects are linked. Here we show that timely removal of PCNA from DNA by the Elg1 complex is important to prevent mutations. Although premature unloading of PCNA generally increases mutation rate, the mutator phenotype of elg1Δ is attenuated by PCNA mutants PCNA-R14E and PCNA-D150E that spontaneously fall off DNA. In contrast, the elg1Δ mutator phenotype is exacerbated by PCNA mutants that accumulate on DNA due to enhanced electrostatic PCNA–DNA interactions. Epistasis analysis suggests that PCNA over-accumulation on DNA interferes with both MMR and MMR-independent process(es). In elg1Δ, over-retained PCNA hyper-recruits the Msh2–Msh6 mismatch recognition complex through its PCNA-interacting peptide motif, causing accumulation of MMR intermediates. Our results suggest that PCNA retention controlled by the Elg1 complex is critical for efficient MMR: PCNA needs to be on DNA long enough to enable MMR, but if it is retained too long it interferes with downstream repair steps.

AB - Proliferating cell nuclear antigen (PCNA) is a sliding clamp that acts as a central co-ordinator for mismatch repair (MMR) as well as DNA replication. Loss of Elg1, the major subunit of the PCNA unloader complex, causes over-accumulation of PCNA on DNA and also increases mutation rate, but it has been unclear if the two effects are linked. Here we show that timely removal of PCNA from DNA by the Elg1 complex is important to prevent mutations. Although premature unloading of PCNA generally increases mutation rate, the mutator phenotype of elg1Δ is attenuated by PCNA mutants PCNA-R14E and PCNA-D150E that spontaneously fall off DNA. In contrast, the elg1Δ mutator phenotype is exacerbated by PCNA mutants that accumulate on DNA due to enhanced electrostatic PCNA–DNA interactions. Epistasis analysis suggests that PCNA over-accumulation on DNA interferes with both MMR and MMR-independent process(es). In elg1Δ, over-retained PCNA hyper-recruits the Msh2–Msh6 mismatch recognition complex through its PCNA-interacting peptide motif, causing accumulation of MMR intermediates. Our results suggest that PCNA retention controlled by the Elg1 complex is critical for efficient MMR: PCNA needs to be on DNA long enough to enable MMR, but if it is retained too long it interferes with downstream repair steps.

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KW - CELL NUCLEAR ANTIGEN

KW - SACCHAROMYCES-CEREVISIAE

KW - GENOME STABILITY

KW - S-PHASE

KW - TRANSLESION SYNTHESIS

KW - TELOMERE LENGTH

KW - PROTEIN

KW - FORMS

KW - INSTABILITY

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DO - 10.1093/nar/gkz441

M3 - Article

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SN - 0305-1048

VL - 47

SP - 6826

EP - 6841

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 13

ER -

Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex

Abstract

Bibliographical note

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Takashi Kubota

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Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex

Abstract

Bibliographical note

Keywords

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Takashi Kubota

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