Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence

Detalhes bibliográficos
Autor(a) principal: Barros, Andrea C. de [UNESP]
Data de Publicação: 2016
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.jmb.2016.01.019
http://hdl.handle.net/11449/161646
Resumo: Xeroderma pigmentosum type G (XPG) proteins are involved in DNA lesion recognition and promotion of nucleotide excision repair. Specific mutations in these proteins may lead to Cockayne syndrome, in which the patients may display severe developmental retardation and neurological abnormalities. No structural information is available for their spacer region or the C-terminal domain, which are important, respectively, for specific nucleotide excision repair activity and substrate specificity, as well as nuclear translocation. Immunofluorescence studies suggested two specific regions of the XPG C-terminus as potential bipartite nuclear localization sequences, which would be responsible for its translocation to the nucleus by the classical nuclear import pathway mediated by the importin-alpha (Imp alpha). Thus, in order to test these hypotheses and gain insight into the structural basis for the nuclear import process for the XPG protein, we solved the crystal structures of complexes formed by the Imp alpha and peptides corresponding to both putative nuclear localization signal (NLS) sequences (XPG1 and XPG2) and performed isothermal titration calorimetry assays to determine their binding affinities. Structural experiments confirm the binding of both NLS peptides to Impa but, unexpectedly, they bind to the receptor as monopartite NLSs. The isothermal titration calorimetry assays demonstrated that XPG1 and XPG2 peptides bind to two separate binding sites, but with high affinity to the major NLS-binding site of the Imp alpha, resembling classical monopartite SV40 TAg NLS. The results lead to insights about what distinguishes monopartite and bipartite NLSs, as well as the differential roles of XPG1 and XPG2 NLSs in the nuclear localization of XPG. (c) 2016 Elsevier Ltd.
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spelling Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS SequenceXeroderma pigmentosum type G (XPG) proteins are involved in DNA lesion recognition and promotion of nucleotide excision repair. Specific mutations in these proteins may lead to Cockayne syndrome, in which the patients may display severe developmental retardation and neurological abnormalities. No structural information is available for their spacer region or the C-terminal domain, which are important, respectively, for specific nucleotide excision repair activity and substrate specificity, as well as nuclear translocation. Immunofluorescence studies suggested two specific regions of the XPG C-terminus as potential bipartite nuclear localization sequences, which would be responsible for its translocation to the nucleus by the classical nuclear import pathway mediated by the importin-alpha (Imp alpha). Thus, in order to test these hypotheses and gain insight into the structural basis for the nuclear import process for the XPG protein, we solved the crystal structures of complexes formed by the Imp alpha and peptides corresponding to both putative nuclear localization signal (NLS) sequences (XPG1 and XPG2) and performed isothermal titration calorimetry assays to determine their binding affinities. Structural experiments confirm the binding of both NLS peptides to Impa but, unexpectedly, they bind to the receptor as monopartite NLSs. The isothermal titration calorimetry assays demonstrated that XPG1 and XPG2 peptides bind to two separate binding sites, but with high affinity to the major NLS-binding site of the Imp alpha, resembling classical monopartite SV40 TAg NLS. The results lead to insights about what distinguishes monopartite and bipartite NLSs, as well as the differential roles of XPG1 and XPG2 NLSs in the nuclear localization of XPG. (c) 2016 Elsevier Ltd.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-18618970 Botucatu, SP, BrazilUniv Zaragoza, Inst Biocomputat & Phys Complex Syst, Consejo Super Invest Cient, Inst Biocomputat & Phys Complex Syst,Joint Unit I, Zaragoza 50018, SpainUniv Zaragoza, Dept Biochem & Mol & Cell Biol, Zaragoza 50018, SpainGovt Aragon, Fdn Agencia Aragonesa Invest & Desarrollo, Zaragoza 50018, 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-18618970 Botucatu, SP, BrazilSpanish Ministerio de Economia y Competitividad: BFU2013-47064-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]2018-11-26T16:48:05Z2018-11-26T16:48:05Z2016-05-22info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article2120-2131application/pdfhttp://dx.doi.org/10.1016/j.jmb.2016.01.019Journal Of Molecular Biology. London: Academic Press Ltd- Elsevier Science Ltd, v. 428, n. 10, p. 2120-2131, 2016.0022-2836http://hdl.handle.net/11449/16164610.1016/j.jmb.2016.01.019WOS:000378444800011WOS000378444800011.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal Of Molecular Biology3,393info:eu-repo/semantics/openAccess2023-11-06T06:06:22Zoai:repositorio.unesp.br:11449/161646Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T17:00:07.778762Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
title Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
spellingShingle Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
Barros, Andrea C. de [UNESP]
title_short Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
title_full Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
title_fullStr Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
title_full_unstemmed Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
title_sort Structural and Calorimetric Studies Demonstrate that Xeroderma Pigmentosum Type G (XPG) Can Be Imported to the Nucleus by a Classical Nuclear Import Pathway via a Monopartite NLS Sequence
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]
description Xeroderma pigmentosum type G (XPG) proteins are involved in DNA lesion recognition and promotion of nucleotide excision repair. Specific mutations in these proteins may lead to Cockayne syndrome, in which the patients may display severe developmental retardation and neurological abnormalities. No structural information is available for their spacer region or the C-terminal domain, which are important, respectively, for specific nucleotide excision repair activity and substrate specificity, as well as nuclear translocation. Immunofluorescence studies suggested two specific regions of the XPG C-terminus as potential bipartite nuclear localization sequences, which would be responsible for its translocation to the nucleus by the classical nuclear import pathway mediated by the importin-alpha (Imp alpha). Thus, in order to test these hypotheses and gain insight into the structural basis for the nuclear import process for the XPG protein, we solved the crystal structures of complexes formed by the Imp alpha and peptides corresponding to both putative nuclear localization signal (NLS) sequences (XPG1 and XPG2) and performed isothermal titration calorimetry assays to determine their binding affinities. Structural experiments confirm the binding of both NLS peptides to Impa but, unexpectedly, they bind to the receptor as monopartite NLSs. The isothermal titration calorimetry assays demonstrated that XPG1 and XPG2 peptides bind to two separate binding sites, but with high affinity to the major NLS-binding site of the Imp alpha, resembling classical monopartite SV40 TAg NLS. The results lead to insights about what distinguishes monopartite and bipartite NLSs, as well as the differential roles of XPG1 and XPG2 NLSs in the nuclear localization of XPG. (c) 2016 Elsevier Ltd.
publishDate 2016
dc.date.none.fl_str_mv 2016-05-22
2018-11-26T16:48:05Z
2018-11-26T16:48:05Z
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.jmb.2016.01.019
Journal Of Molecular Biology. London: Academic Press Ltd- Elsevier Science Ltd, v. 428, n. 10, p. 2120-2131, 2016.
0022-2836
http://hdl.handle.net/11449/161646
10.1016/j.jmb.2016.01.019
WOS:000378444800011
WOS000378444800011.pdf
url http://dx.doi.org/10.1016/j.jmb.2016.01.019
http://hdl.handle.net/11449/161646
identifier_str_mv Journal Of Molecular Biology. London: Academic Press Ltd- Elsevier Science Ltd, v. 428, n. 10, p. 2120-2131, 2016.
0022-2836
10.1016/j.jmb.2016.01.019
WOS:000378444800011
WOS000378444800011.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal Of Molecular Biology
3,393
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 2120-2131
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
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instacron_str UNESP
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