DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway

Detalhes bibliográficos
Autor(a) principal: Barros, Andrea C. de [UNESP]
Data de Publicação: 2018
Outros Autores: Takeda, Agnes A. S. [UNESP], Dreyer, Thiago R. [UNESP], Velazquez-Campoy, Adrian, Kobe, Bostjan, Fontes, Marcos R. M. [UNESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1016/j.biochi.2017.11.013
http://hdl.handle.net/11449/163868
Resumo: MLH1 and PMS2 proteins form the MutLa heterodimer, which plays a major role in DNA mismatch repair (MMR) in humans. Mutations in MMR-related proteins are associated with cancer, especially with colon cancer. The N-terminal region of MutLa comprises the N-termini of PMS2 and MLH1 and, similarly, the C-terminal region of MutLa is composed by the C-termini of PMS2 and MLH1, and the two are connected by linker region. The nuclear localization sequences (NLSs) necessary for the nuclear transport of the two proteins are found in this linker region. However, the exact NLS sequences have been controversial, with different sequences reported, particularly for MLH1. The individual components are not imported efficiently, presumably due to their C-termini masking their NLSs. In order to gain insights into the nuclear transport of these proteins, we solved the crystal structures of importin-alpha bound to peptides corresponding to the supposed NLSs of MLH1 and PMS2 and performed isothermal titration calorimetry to study their binding affinities. Both putative MLH1 and PMS2 NLSs can bind to importin-alpha as monopartite NLSs, which is in agreement with some previous studies. However, MLH1-NLS has the highest affinity measured by a natural NLS peptide, suggesting a major role of MLH1 protein in nuclear import compared to PMS2. Finally, the role of MLH1 and PMS2 in the nuclear transport of the MutLa heterodimer is discussed. (C) 2017 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.
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spelling DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathwayImportin-alphaNuclear import pathwayNuclear localization sequence (NLS)DNA repairMLH1 and PMS2 proteinsX-ray crystallographyIsothermal titration calorimetryMLH1 and PMS2 proteins form the MutLa heterodimer, which plays a major role in DNA mismatch repair (MMR) in humans. Mutations in MMR-related proteins are associated with cancer, especially with colon cancer. The N-terminal region of MutLa comprises the N-termini of PMS2 and MLH1 and, similarly, the C-terminal region of MutLa is composed by the C-termini of PMS2 and MLH1, and the two are connected by linker region. The nuclear localization sequences (NLSs) necessary for the nuclear transport of the two proteins are found in this linker region. However, the exact NLS sequences have been controversial, with different sequences reported, particularly for MLH1. The individual components are not imported efficiently, presumably due to their C-termini masking their NLSs. In order to gain insights into the nuclear transport of these proteins, we solved the crystal structures of importin-alpha bound to peptides corresponding to the supposed NLSs of MLH1 and PMS2 and performed isothermal titration calorimetry to study their binding affinities. Both putative MLH1 and PMS2 NLSs can bind to importin-alpha as monopartite NLSs, which is in agreement with some previous studies. However, MLH1-NLS has the highest affinity measured by a natural NLS peptide, suggesting a major role of MLH1 protein in nuclear import compared to PMS2. Finally, the role of MLH1 and PMS2 in the nuclear transport of the MutLa heterodimer is discussed. (C) 2017 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Spanish Ministerio de Economia y CompetitividadUniv Estadual Paulista, Inst Biociencias, Dept Fis & Biofis, BR-18618689 Botucatu, SP, BrazilUniv Zaragoza, Joint Unit IQFR CSIC BIFI, Inst Biocomputat & Phys Complex Syst BIFI, Zaragoza, SpainUniv Zaragoza, Dep Biochem & Mol & Cell Biol, Zaragoza, SpainGovt Aragon, Fdn ARAID, Zaragoza, SpainUniv Queensland, Sch Chem & Mol Biosci, Inst Mol Biosci, Brisbane, Qld 4072, AustraliaUniv Queensland, Australian Infect Dis Res Ctr, Brisbane, Qld 4072, AustraliaUniv Estadual Paulista, Inst Biociencias, Dept Fis & Biofis, BR-18618689 Botucatu, SP, BrazilFAPESP: 2013/24705-3FAPESP: 2009/14118-8Spanish Ministerio de Economia y Competitividad: BFU2016-78232-PElsevier B.V.Universidade Estadual Paulista (Unesp)Univ ZaragozaGovt AragonUniv QueenslandBarros, Andrea C. de [UNESP]Takeda, Agnes A. S. [UNESP]Dreyer, Thiago R. [UNESP]Velazquez-Campoy, AdrianKobe, BostjanFontes, Marcos R. M. [UNESP]2018-11-26T17:48:14Z2018-11-26T17:48:14Z2018-03-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article87-96application/pdfhttp://dx.doi.org/10.1016/j.biochi.2017.11.013Biochimie. Issy-les-moulineaux: Elsevier France-editions Scientifiques Medicales Elsevier, v. 146, p. 87-96, 2018.0300-9084http://hdl.handle.net/11449/16386810.1016/j.biochi.2017.11.013WOS:000425283600011WOS000425283600011.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiochimie1,554info:eu-repo/semantics/openAccess2024-01-27T06:59:03Zoai:repositorio.unesp.br:11449/163868Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-06T00:06:08.578113Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
title DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
spellingShingle DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
Barros, Andrea C. de [UNESP]
Importin-alpha
Nuclear import pathway
Nuclear localization sequence (NLS)
DNA repair
MLH1 and PMS2 proteins
X-ray crystallography
Isothermal titration calorimetry
title_short DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
title_full DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
title_fullStr DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
title_full_unstemmed DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
title_sort DNA mismatch repair proteins MLH1 and PMS2 can be imported to the nucleus by a classical nuclear import pathway
author Barros, Andrea C. de [UNESP]
author_facet Barros, Andrea C. de [UNESP]
Takeda, Agnes A. S. [UNESP]
Dreyer, Thiago R. [UNESP]
Velazquez-Campoy, Adrian
Kobe, Bostjan
Fontes, Marcos R. M. [UNESP]
author_role author
author2 Takeda, Agnes A. S. [UNESP]
Dreyer, Thiago R. [UNESP]
Velazquez-Campoy, Adrian
Kobe, Bostjan
Fontes, Marcos R. M. [UNESP]
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Univ Zaragoza
Govt Aragon
Univ Queensland
dc.contributor.author.fl_str_mv Barros, Andrea C. de [UNESP]
Takeda, Agnes A. S. [UNESP]
Dreyer, Thiago R. [UNESP]
Velazquez-Campoy, Adrian
Kobe, Bostjan
Fontes, Marcos R. M. [UNESP]
dc.subject.por.fl_str_mv Importin-alpha
Nuclear import pathway
Nuclear localization sequence (NLS)
DNA repair
MLH1 and PMS2 proteins
X-ray crystallography
Isothermal titration calorimetry
topic Importin-alpha
Nuclear import pathway
Nuclear localization sequence (NLS)
DNA repair
MLH1 and PMS2 proteins
X-ray crystallography
Isothermal titration calorimetry
description MLH1 and PMS2 proteins form the MutLa heterodimer, which plays a major role in DNA mismatch repair (MMR) in humans. Mutations in MMR-related proteins are associated with cancer, especially with colon cancer. The N-terminal region of MutLa comprises the N-termini of PMS2 and MLH1 and, similarly, the C-terminal region of MutLa is composed by the C-termini of PMS2 and MLH1, and the two are connected by linker region. The nuclear localization sequences (NLSs) necessary for the nuclear transport of the two proteins are found in this linker region. However, the exact NLS sequences have been controversial, with different sequences reported, particularly for MLH1. The individual components are not imported efficiently, presumably due to their C-termini masking their NLSs. In order to gain insights into the nuclear transport of these proteins, we solved the crystal structures of importin-alpha bound to peptides corresponding to the supposed NLSs of MLH1 and PMS2 and performed isothermal titration calorimetry to study their binding affinities. Both putative MLH1 and PMS2 NLSs can bind to importin-alpha as monopartite NLSs, which is in agreement with some previous studies. However, MLH1-NLS has the highest affinity measured by a natural NLS peptide, suggesting a major role of MLH1 protein in nuclear import compared to PMS2. Finally, the role of MLH1 and PMS2 in the nuclear transport of the MutLa heterodimer is discussed. (C) 2017 Elsevier B.V. and Societe Francaise de Biochimie et Biologie Moleculaire (SFBBM). All rights reserved.
publishDate 2018
dc.date.none.fl_str_mv 2018-11-26T17:48:14Z
2018-11-26T17:48:14Z
2018-03-01
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1016/j.biochi.2017.11.013
Biochimie. Issy-les-moulineaux: Elsevier France-editions Scientifiques Medicales Elsevier, v. 146, p. 87-96, 2018.
0300-9084
http://hdl.handle.net/11449/163868
10.1016/j.biochi.2017.11.013
WOS:000425283600011
WOS000425283600011.pdf
url http://dx.doi.org/10.1016/j.biochi.2017.11.013
http://hdl.handle.net/11449/163868
identifier_str_mv Biochimie. Issy-les-moulineaux: Elsevier France-editions Scientifiques Medicales Elsevier, v. 146, p. 87-96, 2018.
0300-9084
10.1016/j.biochi.2017.11.013
WOS:000425283600011
WOS000425283600011.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biochimie
1,554
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 87-96
application/pdf
dc.publisher.none.fl_str_mv Elsevier B.V.
publisher.none.fl_str_mv Elsevier B.V.
dc.source.none.fl_str_mv Web of Science
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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