Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits
Autor(a) principal: | |
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Data de Publicação: | 2002 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/27002 http://dx.doi.org/10.1042/BJ20020556 |
Resumo: | The heterotrimeric eukaryotic initiation factor (eIF) 2 binds the initiator methionyl-tRNA in a GTP-dependent mode and delivers it to the 40 S ribosomal subunit. in the present study, we have identified amino acid residues in eIF2beta required for binding to eIF2gamma in yeast. Alteration of six residues in the central region of eIF2beta abolished this interaction, as determined by GST-pull down and two-hybrid assays, and leads to cell lethality. Substitution of (131)Tyr and (132)Ser by alanine residues ((YS)-Y-131), although abolishing the binding to eIF2gamma in these assays, resulted in a functional but defective protein in vivo, imparting a temperature-sensitive growth phenotype to cells. A dramatically weakened association of this mutant protein with eIF2gamma in vivo was shown by co-immunoprecipitation. the (YS)-Y-131 mutation in eIF2beta allows translation to initiate at non-AUG codons, as defined by the ability of cells carrying an initiator codon mutation in the HIS4 mRNA to grow in the absence of histidine. the combination of this mutation with the (254)Ser --> Tyr alteration, a previously isolated suppressor of initiator codon mutations which has been shown to increase the spontaneous GTP hydrolysis in the ternary complex, caused a recessive lethality, suggesting additive defects. Thus the impaired interaction of these two subunits represents a novel type of defect in eIF2 function, providing in vivo evidence that the strength of interaction between eIF2beta and eIF2gamma defines the correct usage of the AUG codon for translation initiation. |
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Hashimoto, Nilce Naomi [UNIFESP]Carnevalli, Larissa de Souza [UNIFESP]Castilho, Beatriz Amaral de [UNIFESP]Universidade Federal de São Paulo (UNIFESP)2016-01-24T12:33:33Z2016-01-24T12:33:33Z2002-10-15Biochemical Journal. London: Portland Press, v. 367, p. 359-368, 2002.0264-6021http://repositorio.unifesp.br/handle/11600/27002http://dx.doi.org/10.1042/BJ2002055610.1042/BJ20020556WOS:000178778800006The heterotrimeric eukaryotic initiation factor (eIF) 2 binds the initiator methionyl-tRNA in a GTP-dependent mode and delivers it to the 40 S ribosomal subunit. in the present study, we have identified amino acid residues in eIF2beta required for binding to eIF2gamma in yeast. Alteration of six residues in the central region of eIF2beta abolished this interaction, as determined by GST-pull down and two-hybrid assays, and leads to cell lethality. Substitution of (131)Tyr and (132)Ser by alanine residues ((YS)-Y-131), although abolishing the binding to eIF2gamma in these assays, resulted in a functional but defective protein in vivo, imparting a temperature-sensitive growth phenotype to cells. A dramatically weakened association of this mutant protein with eIF2gamma in vivo was shown by co-immunoprecipitation. the (YS)-Y-131 mutation in eIF2beta allows translation to initiate at non-AUG codons, as defined by the ability of cells carrying an initiator codon mutation in the HIS4 mRNA to grow in the absence of histidine. the combination of this mutation with the (254)Ser --> Tyr alteration, a previously isolated suppressor of initiator codon mutations which has been shown to increase the spontaneous GTP hydrolysis in the ternary complex, caused a recessive lethality, suggesting additive defects. Thus the impaired interaction of these two subunits represents a novel type of defect in eIF2 function, providing in vivo evidence that the strength of interaction between eIF2beta and eIF2gamma defines the correct usage of the AUG codon for translation initiation.Universidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilWeb of Science359-368engPortland PressBiochemical JournaleIF2 beta mutationsnon-AUG recognitionsubunit interaction determinantstranslation initiationTranslation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunitsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP11600/270022022-07-08 10:58:17.493metadata only accessoai:repositorio.unifesp.br:11600/27002Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652022-07-08T13:58:17Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
title |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
spellingShingle |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits Hashimoto, Nilce Naomi [UNIFESP] eIF2 beta mutations non-AUG recognition subunit interaction determinants translation initiation |
title_short |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
title_full |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
title_fullStr |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
title_full_unstemmed |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
title_sort |
Translation initiation at non-AUG codons mediated by weakened association of eukaryotic initiation factor (eIF) 2 subunits |
author |
Hashimoto, Nilce Naomi [UNIFESP] |
author_facet |
Hashimoto, Nilce Naomi [UNIFESP] Carnevalli, Larissa de Souza [UNIFESP] Castilho, Beatriz Amaral de [UNIFESP] |
author_role |
author |
author2 |
Carnevalli, Larissa de Souza [UNIFESP] Castilho, Beatriz Amaral de [UNIFESP] |
author2_role |
author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Hashimoto, Nilce Naomi [UNIFESP] Carnevalli, Larissa de Souza [UNIFESP] Castilho, Beatriz Amaral de [UNIFESP] |
dc.subject.eng.fl_str_mv |
eIF2 beta mutations non-AUG recognition subunit interaction determinants translation initiation |
topic |
eIF2 beta mutations non-AUG recognition subunit interaction determinants translation initiation |
description |
The heterotrimeric eukaryotic initiation factor (eIF) 2 binds the initiator methionyl-tRNA in a GTP-dependent mode and delivers it to the 40 S ribosomal subunit. in the present study, we have identified amino acid residues in eIF2beta required for binding to eIF2gamma in yeast. Alteration of six residues in the central region of eIF2beta abolished this interaction, as determined by GST-pull down and two-hybrid assays, and leads to cell lethality. Substitution of (131)Tyr and (132)Ser by alanine residues ((YS)-Y-131), although abolishing the binding to eIF2gamma in these assays, resulted in a functional but defective protein in vivo, imparting a temperature-sensitive growth phenotype to cells. A dramatically weakened association of this mutant protein with eIF2gamma in vivo was shown by co-immunoprecipitation. the (YS)-Y-131 mutation in eIF2beta allows translation to initiate at non-AUG codons, as defined by the ability of cells carrying an initiator codon mutation in the HIS4 mRNA to grow in the absence of histidine. the combination of this mutation with the (254)Ser --> Tyr alteration, a previously isolated suppressor of initiator codon mutations which has been shown to increase the spontaneous GTP hydrolysis in the ternary complex, caused a recessive lethality, suggesting additive defects. Thus the impaired interaction of these two subunits represents a novel type of defect in eIF2 function, providing in vivo evidence that the strength of interaction between eIF2beta and eIF2gamma defines the correct usage of the AUG codon for translation initiation. |
publishDate |
2002 |
dc.date.issued.fl_str_mv |
2002-10-15 |
dc.date.accessioned.fl_str_mv |
2016-01-24T12:33:33Z |
dc.date.available.fl_str_mv |
2016-01-24T12:33:33Z |
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.citation.fl_str_mv |
Biochemical Journal. London: Portland Press, v. 367, p. 359-368, 2002. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/27002 http://dx.doi.org/10.1042/BJ20020556 |
dc.identifier.issn.none.fl_str_mv |
0264-6021 |
dc.identifier.doi.none.fl_str_mv |
10.1042/BJ20020556 |
dc.identifier.wos.none.fl_str_mv |
WOS:000178778800006 |
identifier_str_mv |
Biochemical Journal. London: Portland Press, v. 367, p. 359-368, 2002. 0264-6021 10.1042/BJ20020556 WOS:000178778800006 |
url |
http://repositorio.unifesp.br/handle/11600/27002 http://dx.doi.org/10.1042/BJ20020556 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Biochemical Journal |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
359-368 |
dc.publisher.none.fl_str_mv |
Portland Press |
publisher.none.fl_str_mv |
Portland Press |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
instname_str |
Universidade Federal de São Paulo (UNIFESP) |
instacron_str |
UNIFESP |
institution |
UNIFESP |
reponame_str |
Repositório Institucional da UNIFESP |
collection |
Repositório Institucional da UNIFESP |
repository.name.fl_str_mv |
Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP) |
repository.mail.fl_str_mv |
|
_version_ |
1802764157500522496 |