Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification
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.1742-4658.2007.06172.x http://hdl.handle.net/11449/41225 |
Resumo: | The eukaryotic translation initiation factor 5A (eIF5A) is the only protein that contains hypusine [N-epsilon-(4-amino-2-hydroxybutyl)lysine], which is required for its activity. Hypusine is formed by post-translational modification of one specific lysine (Lys50 for human eIF5A) by deoxyhypusine synthase and deoxyhypusine hydroxylase. To investigate the features of eIF5A required for its activity, we generated 49 mutations in human eIF5A-1, with a single amino acid substitution at the highly conserved residues or with N-terminal or C-terminal truncations, and tested mutant proteins in complementing the growth of a Saccharomyces cerevisiae eIF5A null strain. Growth-supporting activity was abolished in only a few mutant eIF5As (K47D, G49A, K50A, K50D, K50I, K50R, G52A and K55A), with substitutions at or near the hypusine modification site or with truncation of 21 amino acids from either the N-terminus or C-terminus. The inactivity of the Lys50 substitution proteins is obviously due to lack of deoxyhypusine modification. In contrast, K47D and G49A were effective substrates for deoxyhypusine synthase, yet failed to support growth, suggesting critical roles of Lys47 and Gly49 in eIF5A activity, possibly in its interaction with effector(s). By use of a UBHY-R strain harboring genetically engineered unstable eIF5A, we present evidence for the primary function of eIF5A in protein synthesis. When selected eIF5A mutant proteins were tested for their activity in protein synthesis, a close correlation was observed between their ability to enhance protein synthesis and growth, lending further support for a central role of eIF5A in translation. |
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Repositório Institucional da UNESP |
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Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modificationdeoxyhypusine synthaseeIF5Ahypusinepost-translational modificationtranslation initiationThe eukaryotic translation initiation factor 5A (eIF5A) is the only protein that contains hypusine [N-epsilon-(4-amino-2-hydroxybutyl)lysine], which is required for its activity. Hypusine is formed by post-translational modification of one specific lysine (Lys50 for human eIF5A) by deoxyhypusine synthase and deoxyhypusine hydroxylase. To investigate the features of eIF5A required for its activity, we generated 49 mutations in human eIF5A-1, with a single amino acid substitution at the highly conserved residues or with N-terminal or C-terminal truncations, and tested mutant proteins in complementing the growth of a Saccharomyces cerevisiae eIF5A null strain. Growth-supporting activity was abolished in only a few mutant eIF5As (K47D, G49A, K50A, K50D, K50I, K50R, G52A and K55A), with substitutions at or near the hypusine modification site or with truncation of 21 amino acids from either the N-terminus or C-terminus. The inactivity of the Lys50 substitution proteins is obviously due to lack of deoxyhypusine modification. In contrast, K47D and G49A were effective substrates for deoxyhypusine synthase, yet failed to support growth, suggesting critical roles of Lys47 and Gly49 in eIF5A activity, possibly in its interaction with effector(s). By use of a UBHY-R strain harboring genetically engineered unstable eIF5A, we present evidence for the primary function of eIF5A in protein synthesis. When selected eIF5A mutant proteins were tested for their activity in protein synthesis, a close correlation was observed between their ability to enhance protein synthesis and growth, lending further support for a central role of eIF5A in translation.NIDCR, OPCB, NIH, Bethesda, MD 20892 USASão Paulo State Univ, Sch Pharmaceut Sci, Dept Biol Sci, São Paulo, BrazilBiosearch Technol Inc, Novato, CA USAUniv Calif Davis, Sch Med, Dept Biochem & Mol Med, Davis, CA 95616 USASão Paulo State Univ, Sch Pharmaceut Sci, Dept Biol Sci, São Paulo, BrazilBlackwell PublishingNIDCRUniversidade Estadual Paulista (Unesp)Biosearch Technol IncUniv Calif DavisCano, Veridiana S. P. [UNESP]Jeon, Geoung A.Johansson, Hans E.Henderson, C. AllenPark, Jong-HwanValentini, Sandro Roberto [UNESP]Hershey, John W. B.Park, Myung Hee2014-05-20T15:32:17Z2014-05-20T15:32:17Z2008-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article44-58http://dx.doi.org/10.1111/j.1742-4658.2007.06172.xFebs Journal. Oxford: Blackwell Publishing, v. 275, n. 1, p. 44-58, 2008.1742-464Xhttp://hdl.handle.net/11449/4122510.1111/j.1742-4658.2007.06172.xWOS:0002517648000045333250355049814Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengFEBS Journal4.530info:eu-repo/semantics/openAccess2021-10-23T02:05:39Zoai:repositorio.unesp.br:11449/41225Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T02:05:39Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
title |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
spellingShingle |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification Cano, Veridiana S. P. [UNESP] deoxyhypusine synthase eIF5A hypusine post-translational modification translation initiation |
title_short |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
title_full |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
title_fullStr |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
title_full_unstemmed |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
title_sort |
Mutational analyses of human eIF5A-1-identification of amino acid residues critical for eIF5A activity and hypusine modification |
author |
Cano, Veridiana S. P. [UNESP] |
author_facet |
Cano, Veridiana S. P. [UNESP] Jeon, Geoung A. Johansson, Hans E. Henderson, C. Allen Park, Jong-Hwan Valentini, Sandro Roberto [UNESP] Hershey, John W. B. Park, Myung Hee |
author_role |
author |
author2 |
Jeon, Geoung A. Johansson, Hans E. Henderson, C. Allen Park, Jong-Hwan Valentini, Sandro Roberto [UNESP] Hershey, John W. B. Park, Myung Hee |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
NIDCR Universidade Estadual Paulista (Unesp) Biosearch Technol Inc Univ Calif Davis |
dc.contributor.author.fl_str_mv |
Cano, Veridiana S. P. [UNESP] Jeon, Geoung A. Johansson, Hans E. Henderson, C. Allen Park, Jong-Hwan Valentini, Sandro Roberto [UNESP] Hershey, John W. B. Park, Myung Hee |
dc.subject.por.fl_str_mv |
deoxyhypusine synthase eIF5A hypusine post-translational modification translation initiation |
topic |
deoxyhypusine synthase eIF5A hypusine post-translational modification translation initiation |
description |
The eukaryotic translation initiation factor 5A (eIF5A) is the only protein that contains hypusine [N-epsilon-(4-amino-2-hydroxybutyl)lysine], which is required for its activity. Hypusine is formed by post-translational modification of one specific lysine (Lys50 for human eIF5A) by deoxyhypusine synthase and deoxyhypusine hydroxylase. To investigate the features of eIF5A required for its activity, we generated 49 mutations in human eIF5A-1, with a single amino acid substitution at the highly conserved residues or with N-terminal or C-terminal truncations, and tested mutant proteins in complementing the growth of a Saccharomyces cerevisiae eIF5A null strain. Growth-supporting activity was abolished in only a few mutant eIF5As (K47D, G49A, K50A, K50D, K50I, K50R, G52A and K55A), with substitutions at or near the hypusine modification site or with truncation of 21 amino acids from either the N-terminus or C-terminus. The inactivity of the Lys50 substitution proteins is obviously due to lack of deoxyhypusine modification. In contrast, K47D and G49A were effective substrates for deoxyhypusine synthase, yet failed to support growth, suggesting critical roles of Lys47 and Gly49 in eIF5A activity, possibly in its interaction with effector(s). By use of a UBHY-R strain harboring genetically engineered unstable eIF5A, we present evidence for the primary function of eIF5A in protein synthesis. When selected eIF5A mutant proteins were tested for their activity in protein synthesis, a close correlation was observed between their ability to enhance protein synthesis and growth, lending further support for a central role of eIF5A in translation. |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008-01-01 2014-05-20T15:32:17Z 2014-05-20T15:32:17Z |
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.1742-4658.2007.06172.x Febs Journal. Oxford: Blackwell Publishing, v. 275, n. 1, p. 44-58, 2008. 1742-464X http://hdl.handle.net/11449/41225 10.1111/j.1742-4658.2007.06172.x WOS:000251764800004 5333250355049814 |
url |
http://dx.doi.org/10.1111/j.1742-4658.2007.06172.x http://hdl.handle.net/11449/41225 |
identifier_str_mv |
Febs Journal. Oxford: Blackwell Publishing, v. 275, n. 1, p. 44-58, 2008. 1742-464X 10.1111/j.1742-4658.2007.06172.x WOS:000251764800004 5333250355049814 |
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 |
44-58 |
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 |
|
_version_ |
1799965314928082944 |