Unexpected moves: a conformational change in MutSα enables high-affinity DNA mismatch binding

Susanne R Bruekner; Wietske Pieters; Alexander Fish; A Manuel Liaci; Serge Scheffers; Emily Rayner; Daphne Kaldenbach; Lisa Drost; Marleen Dekker; Sandrine van Hees-Stuivenberg; Elly Delzenne-Goette; Charlotte de Konink; Hellen Houlleberghs; Hendrikus Jan Dubbink; Abeer AlSaegh; Niels de Wind; Friedrich Förster; Hein Te Riele; Titia K Sixma

doi:10.1093/nar/gkad015

Unexpected moves: a conformational change in MutSα enables high-affinity DNA mismatch binding

Susanne R Bruekner, Wietske Pieters, Alexander Fish, A Manuel Liaci, Serge Scheffers, Emily Rayner, Daphne Kaldenbach, Lisa Drost, Marleen Dekker, Sandrine van Hees-Stuivenberg, Elly Delzenne-Goette, Charlotte de Konink, Hellen Houlleberghs, Hendrikus Jan Dubbink, Abeer AlSaegh, Niels de Wind, Friedrich Förster, Hein Te Riele^*, Titia K Sixma^*

^*Corresponding author for this work

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

Abstract

The DNA mismatch repair protein MutSα recognizes wrongly incorporated DNA bases and initiates their correction during DNA replication. Dysfunctions in mismatch repair lead to a predisposition to cancer. Here, we study the homozygous mutation V63E in MSH2 that was found in the germline of a patient with suspected constitutional mismatch repair deficiency syndrome who developed colorectal cancer before the age of 30. Characterization of the mutant in mouse models, as well as slippage and repair assays, shows a mildly pathogenic phenotype. Using cryogenic electron microscopy and surface plasmon resonance, we explored the mechanistic effect of this mutation on MutSα function. We discovered that V63E disrupts a previously unappreciated interface between the mismatch binding domains (MBDs) of MSH2 and MSH6 and leads to reduced DNA binding. Our research identifies this interface as a 'safety lock' that ensures high-affinity DNA binding to increase replication fidelity. Our mechanistic model explains the hypomorphic phenotype of the V63E patient mutation and other variants in the MBD interface.

Original language	English
Pages (from-to)	1173 - 1188
Number of pages	16
Journal	Nucleic Acids Research
Volume	51
Issue number	3
Early online date	30 Jan 2023
DOIs	https://doi.org/10.1093/nar/gkad015
Publication status	Published - 22 Feb 2023

Bibliographical note

Publisher Copyright:
© 2023 The Author(s). Published by Oxford University Press on behalf of Nucleic Acids Research.

Funding

FUNDING Dutch Research Council [NWO-TOP 714.016.002 to T.K.S]; European Commission [H2020-MSCA-ITN-2016 722433DNAREPAIRMAN to T.K.S]; Dutch Cancer Society [KWF 2014-7176 to H.t.R.,KWF2016-10645 to H.t.R. and N.d.W.]; European Commision [ERC Consolidator grant 724425 to F.F]. Funding for open access charge: Netherlands Cancer Institute.

Funders	Funder number
FUNDING Dutch Research Council
NWO-TOP	714.016.002
Netherlands Cancer Institute
European Commission	H2020-MSCA-ITN-2016 722433DNAREPAIRMAN
European Research Council	724425
KWF Kankerbestrijding	KWF 2014-7176

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1093/nar/gkad015Licence: CC BY-NC

gkad015Final published version, 5.26 MBLicence: CC BY-NC

Cite this

Bruekner, S. R., Pieters, W., Fish, A., Liaci, A. M., Scheffers, S., Rayner, E., Kaldenbach, D., Drost, L., Dekker, M., van Hees-Stuivenberg, S., Delzenne-Goette, E., de Konink, C., Houlleberghs, H., Dubbink, H. J., AlSaegh, A., de Wind, N., Förster, F., Te Riele, H., & Sixma, T. K. (2023). Unexpected moves: a conformational change in MutSα enables high-affinity DNA mismatch binding. Nucleic Acids Research, 51(3), 1173 - 1188. https://doi.org/10.1093/nar/gkad015

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title = "Unexpected moves: a conformational change in MutSα enables high-affinity DNA mismatch binding",

abstract = "The DNA mismatch repair protein MutSα recognizes wrongly incorporated DNA bases and initiates their correction during DNA replication. Dysfunctions in mismatch repair lead to a predisposition to cancer. Here, we study the homozygous mutation V63E in MSH2 that was found in the germline of a patient with suspected constitutional mismatch repair deficiency syndrome who developed colorectal cancer before the age of 30. Characterization of the mutant in mouse models, as well as slippage and repair assays, shows a mildly pathogenic phenotype. Using cryogenic electron microscopy and surface plasmon resonance, we explored the mechanistic effect of this mutation on MutSα function. We discovered that V63E disrupts a previously unappreciated interface between the mismatch binding domains (MBDs) of MSH2 and MSH6 and leads to reduced DNA binding. Our research identifies this interface as a 'safety lock' that ensures high-affinity DNA binding to increase replication fidelity. Our mechanistic model explains the hypomorphic phenotype of the V63E patient mutation and other variants in the MBD interface.",

author = "Bruekner, {Susanne R} and Wietske Pieters and Alexander Fish and Liaci, {A Manuel} and Serge Scheffers and Emily Rayner and Daphne Kaldenbach and Lisa Drost and Marleen Dekker and {van Hees-Stuivenberg}, Sandrine and Elly Delzenne-Goette and {de Konink}, Charlotte and Hellen Houlleberghs and Dubbink, {Hendrikus Jan} and Abeer AlSaegh and {de Wind}, Niels and Friedrich F{\"o}rster and {Te Riele}, Hein and Sixma, {Titia K}",

note = "Publisher Copyright: {\textcopyright} 2023 The Author(s). Published by Oxford University Press on behalf of Nucleic Acids Research.",

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month = feb,

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doi = "10.1093/nar/gkad015",

language = "English",

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Bruekner, SR, Pieters, W, Fish, A, Liaci, AM, Scheffers, S, Rayner, E, Kaldenbach, D, Drost, L, Dekker, M, van Hees-Stuivenberg, S, Delzenne-Goette, E, de Konink, C, Houlleberghs, H, Dubbink, HJ, AlSaegh, A, de Wind, N, Förster, F, Te Riele, H & Sixma, TK 2023, 'Unexpected moves: a conformational change in MutSα enables high-affinity DNA mismatch binding', Nucleic Acids Research, vol. 51, no. 3, pp. 1173 - 1188. https://doi.org/10.1093/nar/gkad015

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T2 - a conformational change in MutSα enables high-affinity DNA mismatch binding

AU - Bruekner, Susanne R

AU - Pieters, Wietske

AU - Fish, Alexander

AU - Liaci, A Manuel

AU - Scheffers, Serge

AU - Rayner, Emily

AU - Kaldenbach, Daphne

AU - Drost, Lisa

AU - Dekker, Marleen

AU - van Hees-Stuivenberg, Sandrine

AU - Delzenne-Goette, Elly

AU - de Konink, Charlotte

AU - Houlleberghs, Hellen

AU - Dubbink, Hendrikus Jan

AU - AlSaegh, Abeer

AU - de Wind, Niels

AU - Förster, Friedrich

AU - Te Riele, Hein

AU - Sixma, Titia K

PY - 2023/2/22

Y1 - 2023/2/22

N2 - The DNA mismatch repair protein MutSα recognizes wrongly incorporated DNA bases and initiates their correction during DNA replication. Dysfunctions in mismatch repair lead to a predisposition to cancer. Here, we study the homozygous mutation V63E in MSH2 that was found in the germline of a patient with suspected constitutional mismatch repair deficiency syndrome who developed colorectal cancer before the age of 30. Characterization of the mutant in mouse models, as well as slippage and repair assays, shows a mildly pathogenic phenotype. Using cryogenic electron microscopy and surface plasmon resonance, we explored the mechanistic effect of this mutation on MutSα function. We discovered that V63E disrupts a previously unappreciated interface between the mismatch binding domains (MBDs) of MSH2 and MSH6 and leads to reduced DNA binding. Our research identifies this interface as a 'safety lock' that ensures high-affinity DNA binding to increase replication fidelity. Our mechanistic model explains the hypomorphic phenotype of the V63E patient mutation and other variants in the MBD interface.

AB - The DNA mismatch repair protein MutSα recognizes wrongly incorporated DNA bases and initiates their correction during DNA replication. Dysfunctions in mismatch repair lead to a predisposition to cancer. Here, we study the homozygous mutation V63E in MSH2 that was found in the germline of a patient with suspected constitutional mismatch repair deficiency syndrome who developed colorectal cancer before the age of 30. Characterization of the mutant in mouse models, as well as slippage and repair assays, shows a mildly pathogenic phenotype. Using cryogenic electron microscopy and surface plasmon resonance, we explored the mechanistic effect of this mutation on MutSα function. We discovered that V63E disrupts a previously unappreciated interface between the mismatch binding domains (MBDs) of MSH2 and MSH6 and leads to reduced DNA binding. Our research identifies this interface as a 'safety lock' that ensures high-affinity DNA binding to increase replication fidelity. Our mechanistic model explains the hypomorphic phenotype of the V63E patient mutation and other variants in the MBD interface.

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DO - 10.1093/nar/gkad015

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SN - 0305-1048

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SP - 1173

EP - 1188

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 3

ER -

Unexpected moves: a conformational change in MutSα enables high-affinity DNA mismatch binding

Abstract

Bibliographical note

Funding

UN SDGs

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