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)

Research output: Contribution to journalArticle

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 languageEnglish
Pages (from-to)6826-6841
Number of pages16
JournalNucleic Acids Research
Volume47
Issue number13
Early online date22 May 2019
DOIs
Publication statusPublished - 26 Jul 2019

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DNA Mismatch Repair
Proliferating Cell Nuclear Antigen
DNA
Mutation Rate
Phenotype
Static Electricity
DNA Replication

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Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex. / Paul Solomon Devakumar, Lovely Jael; Gaubitz, Christl; Lundblad, Victoria; Kelch, Brian A.; Kubota, Takashi (Corresponding Author).

In: Nucleic Acids Research, Vol. 47, No. 13, 26.07.2019, p. 6826-6841.

Research output: Contribution to journalArticle

Paul Solomon Devakumar, Lovely Jael ; Gaubitz, Christl ; Lundblad, Victoria ; Kelch, Brian A. ; Kubota, Takashi. / Effective mismatch repair depends on timely control of PCNA retention on DNA by the Elg1 complex. In: Nucleic Acids Research. 2019 ; Vol. 47, No. 13. pp. 6826-6841.
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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.",
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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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.

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