Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1111/j.1742-4658.2008.06345.x http://hdl.handle.net/11449/7478 |
Resumo: | Eukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal alpha-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins - eIF5A(K56A) and eIF5A(Q22H,L93F)- and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression. |
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Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesiseIF5Ahypusinemutational analysisstructural modelingtranslationEukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal alpha-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins - eIF5A(K56A) and eIF5A(Q22H,L93F)- and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression.Univ Estadual Paulista, Fac Ciencias Farmaceut, Dept Biol Sci, Sch Pharmaceut Sci, BR-14801902 São Paulo, BrazilNatl Inst Dent & Craniofacial Res, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD USAUniv São Paulo, Inst Phys, Dept Phys & Informat, Ctr Struct Mol Biotechnol, BR-05508 São Paulo, BrazilUniv Estadual Paulista, Fac Ciencias Farmaceut, Dept Biol Sci, Sch Pharmaceut Sci, BR-14801902 São Paulo, BrazilBlackwell PublishingUniversidade Estadual Paulista (Unesp)Natl Inst Dent & Craniofacial ResUniversidade de São Paulo (USP)Dias, Camila A. O.Cano, Veridiana S. P.Rangel, Suzana M.Apponi, Luciano H.Frigieri, Mariana C.Muniz, Joao R. C.Garcia, WaniusPark, Myung H.Garratt, Richard C.Zanelli, Cleslei Fernando [UNESP]Valentini, Sandro Roberto [UNESP]2014-05-20T13:24:16Z2014-05-20T13:24:16Z2008-04-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1874-1888http://dx.doi.org/10.1111/j.1742-4658.2008.06345.xFebs Journal. Oxford: Blackwell Publishing, v. 275, n. 8, p. 1874-1888, 2008.1742-464Xhttp://hdl.handle.net/11449/747810.1111/j.1742-4658.2008.06345.xWOS:000254499500025533325035504981415256654089001950000-0001-7831-1149Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengFEBS Journal4.530info:eu-repo/semantics/openAccess2024-06-24T13:07:51Zoai:repositorio.unesp.br:11449/7478Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-06-24T13:07:51Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
title |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
spellingShingle |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis Dias, Camila A. O. eIF5A hypusine mutational analysis structural modeling translation |
title_short |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
title_full |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
title_fullStr |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
title_full_unstemmed |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
title_sort |
Structural modeling and mutational analysis of yeast eukaryotic translation initiation factor 5A reveal new critical residues and reinforce its involvement in protein synthesis |
author |
Dias, Camila A. O. |
author_facet |
Dias, Camila A. O. Cano, Veridiana S. P. Rangel, Suzana M. Apponi, Luciano H. Frigieri, Mariana C. Muniz, Joao R. C. Garcia, Wanius Park, Myung H. Garratt, Richard C. Zanelli, Cleslei Fernando [UNESP] Valentini, Sandro Roberto [UNESP] |
author_role |
author |
author2 |
Cano, Veridiana S. P. Rangel, Suzana M. Apponi, Luciano H. Frigieri, Mariana C. Muniz, Joao R. C. Garcia, Wanius Park, Myung H. Garratt, Richard C. Zanelli, Cleslei Fernando [UNESP] Valentini, Sandro Roberto [UNESP] |
author2_role |
author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) Natl Inst Dent & Craniofacial Res Universidade de São Paulo (USP) |
dc.contributor.author.fl_str_mv |
Dias, Camila A. O. Cano, Veridiana S. P. Rangel, Suzana M. Apponi, Luciano H. Frigieri, Mariana C. Muniz, Joao R. C. Garcia, Wanius Park, Myung H. Garratt, Richard C. Zanelli, Cleslei Fernando [UNESP] Valentini, Sandro Roberto [UNESP] |
dc.subject.por.fl_str_mv |
eIF5A hypusine mutational analysis structural modeling translation |
topic |
eIF5A hypusine mutational analysis structural modeling translation |
description |
Eukaryotic translation initiation factor 5A (eIF5A) is a protein that is highly conserved and essential for cell viability. This factor is the only protein known to contain the unique and essential amino acid residue hypusine. This work focused on the structural and functional characterization of Saccharomyces cerevisiae eIF5A. The tertiary structure of yeast eIF5A was modeled based on the structure of its Leishmania mexicana homologue and this model was used to predict the structural localization of new site-directed and randomly generated mutations. Most of the 40 new mutants exhibited phenotypes that resulted from eIF-5A protein-folding defects. Our data provided evidence that the C-terminal alpha-helix present in yeast eIF5A is an essential structural element, whereas the eIF5A N-terminal 10 amino acid extension not present in archaeal eIF5A homologs, is not. Moreover, the mutants containing substitutions at or in the vicinity of the hypusine modification site displayed nonviable or temperature-sensitive phenotypes and were defective in hypusine modification. Interestingly, two of the temperature-sensitive strains produced stable mutant eIF5A proteins - eIF5A(K56A) and eIF5A(Q22H,L93F)- and showed defects in protein synthesis at the restrictive temperature. Our data revealed important structural features of eIF5A that are required for its vital role in cell viability and underscored an essential function of eIF5A in the translation step of gene expression. |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008-04-01 2014-05-20T13:24:16Z 2014-05-20T13:24:16Z |
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/10.1111/j.1742-4658.2008.06345.x Febs Journal. Oxford: Blackwell Publishing, v. 275, n. 8, p. 1874-1888, 2008. 1742-464X http://hdl.handle.net/11449/7478 10.1111/j.1742-4658.2008.06345.x WOS:000254499500025 5333250355049814 1525665408900195 0000-0001-7831-1149 |
url |
http://dx.doi.org/10.1111/j.1742-4658.2008.06345.x http://hdl.handle.net/11449/7478 |
identifier_str_mv |
Febs Journal. Oxford: Blackwell Publishing, v. 275, n. 8, p. 1874-1888, 2008. 1742-464X 10.1111/j.1742-4658.2008.06345.x WOS:000254499500025 5333250355049814 1525665408900195 0000-0001-7831-1149 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
FEBS Journal 4.530 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
1874-1888 |
dc.publisher.none.fl_str_mv |
Blackwell Publishing |
publisher.none.fl_str_mv |
Blackwell Publishing |
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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1803045393539268608 |