The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics

Mitochondrial single-stranded DNA-binding protein (mtSSB) is essential for mitochondrial DNA (mtDNA) replication. Recently, several mtSSB variants have been associated with autosomal dominant mitochondrial optic atrophy and retinal dystrophy. Here, we have studied at the molecular level the function...

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Autores: Martucci, Martial, Moretton, Amandine, Tarrés-Soler, Aleix, Ropars, Virginie, Lambert, Louise, Vernet, Patrick, Solà, Maria, Falkenberg, Maria, Farge, Geraldine, Wildenberg, Siet van den
Formato: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Recursos:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/375657
Acesso em linha:http://hdl.handle.net/10261/375657
Access Level:acceso abierto
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spelling The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamicsMartucci, MartialMoretton, AmandineTarrés-Soler, AleixRopars, VirginieLambert, LouiseVernet, PatrickSolà, MariaFalkenberg, MariaFarge, GeraldineWildenberg, Siet van denMitochondrial single-stranded DNA-binding protein (mtSSB) is essential for mitochondrial DNA (mtDNA) replication. Recently, several mtSSB variants have been associated with autosomal dominant mitochondrial optic atrophy and retinal dystrophy. Here, we have studied at the molecular level the functional consequences of one of the most severe mtSSB variants, R107Q. We first studied the oligomeric state of this variant and observed that the mtSSBR107Q mutant forms stable tetramers in vitro. On the other hand, we showed, using complementary single-molecule approaches, that mtSSBR107Q displays a lower intramolecular ssDNA compaction ability and a higher ssDNA dissociation rate than the WT protein. Real-time competition experiments for ssDNA-binding showed a marked advantage of mtSSBWT over mtSSBR107Q. Combined, these results show that the R107Q mutation significantly impaired the ssDNA-binding and compacting ability of mtSSB, likely by weakening mtSSB ssDNA wrapping efficiency. These features are in line with our molecular modeling of ssDNA on mtSSB showing that the R107Q mutation may destabilize local interactions and results in an electronegative spot that interrupts an ssDNA-interacting-electropositive patch, thus reducing the potential mtSSB-ssDNA interaction sites.Association Française Contre les Myopathies Téléthon [24167 to G.F., 25052 to A.M.]; Swedish Research Council [2021-00932 to M.F.]; Swedish Cancer Foundation [to M.F.]; Knut and Alice Wallenberg Foundation [2017.0080, 2018.0204 to M.F.]; Spanish Ministry of Science, Innovation and Universities [MCIN/AEI/10.13039/501100011033 to M.S.]; European Regional Development Fund [PID2021-129038NB-I00 to M.S.]; Generalitat de Catalunya [2021_SGR-00425 to M.S.]; the BLI measurements have benefited from the platform PIM of I2BC supported by French Infrastructure for Integrated Structural Biology (FRISBI) [ANR-10-INBS-05].Peer reviewedOxford University PressAssociation Française contre les MyopathiesSwedish Research CouncilKnut and Alice Wallenberg FoundationEuropean CommissionMinisterio de Ciencia, Innovación y Universidades (España)Agencia Estatal de Investigación (España)Generalitat de CatalunyaConsejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202520252024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/375657reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-129038NB-I00The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1093/nar/gkae354https://doi.org/10.1093/nar/gkae354Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3756572026-05-22T06:33:51Z
dc.title.none.fl_str_mv The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
title The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
spellingShingle The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
Martucci, Martial
title_short The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
title_full The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
title_fullStr The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
title_full_unstemmed The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
title_sort The mutation R107Q alters mtSSB ssDNA compaction ability and binding dynamics
dc.creator.none.fl_str_mv Martucci, Martial
Moretton, Amandine
Tarrés-Soler, Aleix
Ropars, Virginie
Lambert, Louise
Vernet, Patrick
Solà, Maria
Falkenberg, Maria
Farge, Geraldine
Wildenberg, Siet van den
author Martucci, Martial
author_facet Martucci, Martial
Moretton, Amandine
Tarrés-Soler, Aleix
Ropars, Virginie
Lambert, Louise
Vernet, Patrick
Solà, Maria
Falkenberg, Maria
Farge, Geraldine
Wildenberg, Siet van den
author_role author
author2 Moretton, Amandine
Tarrés-Soler, Aleix
Ropars, Virginie
Lambert, Louise
Vernet, Patrick
Solà, Maria
Falkenberg, Maria
Farge, Geraldine
Wildenberg, Siet van den
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Association Française contre les Myopathies
Swedish Research Council
Knut and Alice Wallenberg Foundation
European Commission
Ministerio de Ciencia, Innovación y Universidades (España)
Agencia Estatal de Investigación (España)
Generalitat de Catalunya
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
description Mitochondrial single-stranded DNA-binding protein (mtSSB) is essential for mitochondrial DNA (mtDNA) replication. Recently, several mtSSB variants have been associated with autosomal dominant mitochondrial optic atrophy and retinal dystrophy. Here, we have studied at the molecular level the functional consequences of one of the most severe mtSSB variants, R107Q. We first studied the oligomeric state of this variant and observed that the mtSSBR107Q mutant forms stable tetramers in vitro. On the other hand, we showed, using complementary single-molecule approaches, that mtSSBR107Q displays a lower intramolecular ssDNA compaction ability and a higher ssDNA dissociation rate than the WT protein. Real-time competition experiments for ssDNA-binding showed a marked advantage of mtSSBWT over mtSSBR107Q. Combined, these results show that the R107Q mutation significantly impaired the ssDNA-binding and compacting ability of mtSSB, likely by weakening mtSSB ssDNA wrapping efficiency. These features are in line with our molecular modeling of ssDNA on mtSSB showing that the R107Q mutation may destabilize local interactions and results in an electronegative spot that interrupts an ssDNA-interacting-electropositive patch, thus reducing the potential mtSSB-ssDNA interaction sites.
publishDate 2024
dc.date.none.fl_str_mv 2024
2025
2025
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/375657
url http://hdl.handle.net/10261/375657
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-129038NB-I00
The underlying dataset has been published as supplementary material of the article in the publisher platform at DOI https://doi.org/10.1093/nar/gkae354
https://doi.org/10.1093/nar/gkae354
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Oxford University Press
publisher.none.fl_str_mv Oxford University Press
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.811543

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