Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal
Autor(a) principal: | |
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Data de Publicação: | 2015 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0128687 http://hdl.handle.net/11449/128581 |
Resumo: | Neurospora crassa is a filamentous fungus that has been extensively studied as a model organism for eukaryotic biology, providing fundamental insights into cellular processes such as cell signaling, growth and differentiation. To advance in the study of this multicellular organism, an understanding of the specific mechanisms for protein transport into the cell nucleus is essential. Importin-alpha (Imp-alpha) is the receptor for cargo proteins that contain specific nuclear localization signals (NLSs) that play a key role in the classical nuclear import pathway. Structures of Imp-a from different organisms (yeast, rice, mouse, and human) have been determined, revealing that this receptor possesses a conserved structural scaffold. However, recent studies have demonstrated that the Imp alpha mechanism of action may vary significantly for different organisms or for different isoforms from the same organism. Therefore, structural, functional, and biophysical characterization of different Impa proteins is necessary to understand the selectivity of nuclear transport. Here, we determined the first crystal structure of an Impa from a filamentous fungus which is also the highest resolution Impa structure already solved to date (1.75 angstrom). In addition, we performed calorimetric analysis to determine the affinity and thermodynamic parameters of the interaction between Impa and the classical SV40 NLS peptide. The comparison of these data with previous studies on Impa proteins led us to demonstrate that N. crassa Imp-alpha possess specific features that are distinct from mammalian Imp-alpha but exhibit important similarities to rice Imp-alpha, particularly at the minor NLS binding site. |
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Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signalNeurospora crassa is a filamentous fungus that has been extensively studied as a model organism for eukaryotic biology, providing fundamental insights into cellular processes such as cell signaling, growth and differentiation. To advance in the study of this multicellular organism, an understanding of the specific mechanisms for protein transport into the cell nucleus is essential. Importin-alpha (Imp-alpha) is the receptor for cargo proteins that contain specific nuclear localization signals (NLSs) that play a key role in the classical nuclear import pathway. Structures of Imp-a from different organisms (yeast, rice, mouse, and human) have been determined, revealing that this receptor possesses a conserved structural scaffold. However, recent studies have demonstrated that the Imp alpha mechanism of action may vary significantly for different organisms or for different isoforms from the same organism. Therefore, structural, functional, and biophysical characterization of different Impa proteins is necessary to understand the selectivity of nuclear transport. Here, we determined the first crystal structure of an Impa from a filamentous fungus which is also the highest resolution Impa structure already solved to date (1.75 angstrom). In addition, we performed calorimetric analysis to determine the affinity and thermodynamic parameters of the interaction between Impa and the classical SV40 NLS peptide. The comparison of these data with previous studies on Impa proteins led us to demonstrate that N. crassa Imp-alpha possess specific features that are distinct from mammalian Imp-alpha but exhibit important similarities to rice Imp-alpha, particularly at the minor NLS binding site.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Univ Estadual Paulista, UNESP, Inst Biociencias, Dept Fis &Biofis, Botucatu, SP, BrazilUniv Estadual Paulista, UNESP, Inst Quim, Dept Bioquim &Tecnol Quim, Araraquara, SP, BrazilDepartamento de Física e Biofísica, Instituto de Biociências, Universidade Estadual Paulista, UNESP, Botucatu, SP, BrazilDepartamento de Bioquímica e Tecnologia Química, Instituto de Química, Universidade Estadual Paulista, UNESP, Araraquara, SP, BrazilPublic Library ScienceUniversidade Estadual Paulista (Unesp)Bernardes, Natalia E. [UNESP]Takeda, Agnes A. S. [UNESP]Dreyer, Thiago R. [UNESP]Freitas, Fernanda Z. [UNESP]Bertolini, Maria Celia [UNESP]Fontes, Marcos R. M. [UNESP]2015-10-21T13:11:11Z2015-10-21T13:11:11Z2015-06-19info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1-18application/pdfhttp://journals.plos.org/plosone/article?id=10.1371/journal.pone.0128687Plos One. San Francisco: Public Library Science, v. 10, n. 6, p. 1-18, 2015.1932-6203http://hdl.handle.net/11449/12858110.1371/journal.pone.0128687WOS:000356835000027WOS000356835000027.pdf8817669953838863Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPlos One2.7661,164info:eu-repo/semantics/openAccess2021-10-23T21:50:31Zoai:repositorio.unesp.br:11449/128581Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T21:50:31Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
title |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
spellingShingle |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal Bernardes, Natalia E. [UNESP] |
title_short |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
title_full |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
title_fullStr |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
title_full_unstemmed |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
title_sort |
Structure of importin-a from a filamentous fungus in complex with a classical nuclear localization signal |
author |
Bernardes, Natalia E. [UNESP] |
author_facet |
Bernardes, Natalia E. [UNESP] Takeda, Agnes A. S. [UNESP] Dreyer, Thiago R. [UNESP] Freitas, Fernanda Z. [UNESP] Bertolini, Maria Celia [UNESP] Fontes, Marcos R. M. [UNESP] |
author_role |
author |
author2 |
Takeda, Agnes A. S. [UNESP] Dreyer, Thiago R. [UNESP] Freitas, Fernanda Z. [UNESP] Bertolini, Maria Celia [UNESP] Fontes, Marcos R. M. [UNESP] |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Bernardes, Natalia E. [UNESP] Takeda, Agnes A. S. [UNESP] Dreyer, Thiago R. [UNESP] Freitas, Fernanda Z. [UNESP] Bertolini, Maria Celia [UNESP] Fontes, Marcos R. M. [UNESP] |
description |
Neurospora crassa is a filamentous fungus that has been extensively studied as a model organism for eukaryotic biology, providing fundamental insights into cellular processes such as cell signaling, growth and differentiation. To advance in the study of this multicellular organism, an understanding of the specific mechanisms for protein transport into the cell nucleus is essential. Importin-alpha (Imp-alpha) is the receptor for cargo proteins that contain specific nuclear localization signals (NLSs) that play a key role in the classical nuclear import pathway. Structures of Imp-a from different organisms (yeast, rice, mouse, and human) have been determined, revealing that this receptor possesses a conserved structural scaffold. However, recent studies have demonstrated that the Imp alpha mechanism of action may vary significantly for different organisms or for different isoforms from the same organism. Therefore, structural, functional, and biophysical characterization of different Impa proteins is necessary to understand the selectivity of nuclear transport. Here, we determined the first crystal structure of an Impa from a filamentous fungus which is also the highest resolution Impa structure already solved to date (1.75 angstrom). In addition, we performed calorimetric analysis to determine the affinity and thermodynamic parameters of the interaction between Impa and the classical SV40 NLS peptide. The comparison of these data with previous studies on Impa proteins led us to demonstrate that N. crassa Imp-alpha possess specific features that are distinct from mammalian Imp-alpha but exhibit important similarities to rice Imp-alpha, particularly at the minor NLS binding site. |
publishDate |
2015 |
dc.date.none.fl_str_mv |
2015-10-21T13:11:11Z 2015-10-21T13:11:11Z 2015-06-19 |
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://journals.plos.org/plosone/article?id=10.1371/journal.pone.0128687 Plos One. San Francisco: Public Library Science, v. 10, n. 6, p. 1-18, 2015. 1932-6203 http://hdl.handle.net/11449/128581 10.1371/journal.pone.0128687 WOS:000356835000027 WOS000356835000027.pdf 8817669953838863 |
url |
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0128687 http://hdl.handle.net/11449/128581 |
identifier_str_mv |
Plos One. San Francisco: Public Library Science, v. 10, n. 6, p. 1-18, 2015. 1932-6203 10.1371/journal.pone.0128687 WOS:000356835000027 WOS000356835000027.pdf 8817669953838863 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Plos One 2.766 1,164 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
1-18 application/pdf |
dc.publisher.none.fl_str_mv |
Public Library Science |
publisher.none.fl_str_mv |
Public Library Science |
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 |
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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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