The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity
Autor(a) principal: | |
---|---|
Data de Publicação: | 2013 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0060140 http://hdl.handle.net/11449/74994 |
Resumo: | The putative eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein among archaea and eukaryotes that has recently been implicated in the elongation step of translation. eIF5A undergoes an essential and conserved posttranslational modification at a specific lysine to generate the residue hypusine. The enzymes deoxyhypusine synthase (Dys1) and deoxyhypusine hydroxylase (Lia1) catalyze this two-step modification process. Although several Saccharomyces cerevisiae eIF5A mutants have importantly contributed to the study of eIF5A function, no conditional mutant of Dys1 has been described so far. In this study, we generated and characterized the dys1-1 mutant, which showed a strong depletion of mutated Dys1 protein, resulting in more than 2-fold decrease in hypusine levels relative to the wild type. The dys1-1 mutant demonstrated a defect in total protein synthesis, a defect in polysome profile indicative of a translation elongation defect and a reduced association of eIF5A with polysomes. The growth phenotype of dys1-1 mutant is severe, growing only in the presence of 1 M sorbitol, an osmotic stabilizer. Although this phenotype is characteristic of Pkc1 cell wall integrity mutants, the sorbitol requirement from dys1-1 is not associated with cell lysis. We observed that the dys1-1 genetically interacts with the sole yeast protein kinase C (Pkc1) and Asc1, a component of the 40S ribosomal subunit. The dys1-1 mutant was synthetically lethal in combination with asc1Δ and overexpression of TIF51A (eIF5A) or DYS1 is toxic for an asc1Δ strain. Moreover, eIF5A is more associated with translating ribosomes in the absence of Asc1 in the cell. Finally, analysis of the sensitivity to cell wall-perturbing compounds revealed a more similar behavior of the dys1-1 and asc1Δ mutants in comparison with the pkc1Δ mutant. These data suggest a correlated role for eIF5A and Asc1 in coordinating the translational control of a subset of mRNAs associated with cell integrity. © 2013 Galvão et al. |
id |
UNSP_dc797e7a2a5473c80ae3e4dfc21fdcb9 |
---|---|
oai_identifier_str |
oai:repositorio.unesp.br:11449/74994 |
network_acronym_str |
UNSP |
network_name_str |
Repositório Institucional da UNESP |
repository_id_str |
2946 |
spelling |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integritydeoxyhypusine synthasedeoxyhypusine synthase 1deoxyhypusine synthase 1 1fungal proteinhypusineinitiation factor 5Aprotein Asc1protein kinase Csorbitolunclassified drugalleleasc1 genebinding affinitycell growthcell wallcell wall integritycontrolled studycytolysisdeoxyhypusine synthase 1 geneeukaryotic translation initiation factor 5A genegenegene functiongene interactiongene overexpressiongenetic identificationmutantnonhumanosmosispolysomeprotein depletionprotein functionprotein kinase C geneprotein modificationprotein protein interactionprotein synthesisribosomeRNA translationsensitivity analysistranslation regulationwild typeyeastThe putative eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein among archaea and eukaryotes that has recently been implicated in the elongation step of translation. eIF5A undergoes an essential and conserved posttranslational modification at a specific lysine to generate the residue hypusine. The enzymes deoxyhypusine synthase (Dys1) and deoxyhypusine hydroxylase (Lia1) catalyze this two-step modification process. Although several Saccharomyces cerevisiae eIF5A mutants have importantly contributed to the study of eIF5A function, no conditional mutant of Dys1 has been described so far. In this study, we generated and characterized the dys1-1 mutant, which showed a strong depletion of mutated Dys1 protein, resulting in more than 2-fold decrease in hypusine levels relative to the wild type. The dys1-1 mutant demonstrated a defect in total protein synthesis, a defect in polysome profile indicative of a translation elongation defect and a reduced association of eIF5A with polysomes. The growth phenotype of dys1-1 mutant is severe, growing only in the presence of 1 M sorbitol, an osmotic stabilizer. Although this phenotype is characteristic of Pkc1 cell wall integrity mutants, the sorbitol requirement from dys1-1 is not associated with cell lysis. We observed that the dys1-1 genetically interacts with the sole yeast protein kinase C (Pkc1) and Asc1, a component of the 40S ribosomal subunit. The dys1-1 mutant was synthetically lethal in combination with asc1Δ and overexpression of TIF51A (eIF5A) or DYS1 is toxic for an asc1Δ strain. Moreover, eIF5A is more associated with translating ribosomes in the absence of Asc1 in the cell. Finally, analysis of the sensitivity to cell wall-perturbing compounds revealed a more similar behavior of the dys1-1 and asc1Δ mutants in comparison with the pkc1Δ mutant. These data suggest a correlated role for eIF5A and Asc1 in coordinating the translational control of a subset of mRNAs associated with cell integrity. © 2013 Galvão et al.Department of Biological Sciences Univ Estadual Paulista - UNESP, Araraquara-Saõ PauloDepartment of Biological Sciences Univ Estadual Paulista - UNESP, Araraquara-Saõ PauloUniversidade Estadual Paulista (Unesp)Galvão, Fabio Carrilho [UNESP]Rossi, Danuza [UNESP]Silveira, Wagner da Silva [UNESP]Valentini, Sandro Roberto [UNESP]Zanelli, Cleslei Fernando [UNESP]2014-05-27T11:28:48Z2014-05-27T11:28:48Z2013-04-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1371/journal.pone.0060140PLoS ONE, v. 8, n. 4, 2013.1932-6203http://hdl.handle.net/11449/7499410.1371/journal.pone.0060140WOS:0003169309000502-s2.0-848756864702-s2.0-84875686470.pdf533325035504981415256654089001950000-0001-7831-1149Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPLOS ONE2.7661,164info:eu-repo/semantics/openAccess2024-01-21T06:23:48Zoai:repositorio.unesp.br:11449/74994Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-01-21T06:23:48Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
title |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
spellingShingle |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity Galvão, Fabio Carrilho [UNESP] deoxyhypusine synthase deoxyhypusine synthase 1 deoxyhypusine synthase 1 1 fungal protein hypusine initiation factor 5A protein Asc1 protein kinase C sorbitol unclassified drug allele asc1 gene binding affinity cell growth cell wall cell wall integrity controlled study cytolysis deoxyhypusine synthase 1 gene eukaryotic translation initiation factor 5A gene gene gene function gene interaction gene overexpression genetic identification mutant nonhuman osmosis polysome protein depletion protein function protein kinase C gene protein modification protein protein interaction protein synthesis ribosome RNA translation sensitivity analysis translation regulation wild type yeast |
title_short |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
title_full |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
title_fullStr |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
title_full_unstemmed |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
title_sort |
The Deoxyhypusine Synthase Mutant dys1-1 Reveals the Association of eIF5A and Asc1 with Cell Wall Integrity |
author |
Galvão, Fabio Carrilho [UNESP] |
author_facet |
Galvão, Fabio Carrilho [UNESP] Rossi, Danuza [UNESP] Silveira, Wagner da Silva [UNESP] Valentini, Sandro Roberto [UNESP] Zanelli, Cleslei Fernando [UNESP] |
author_role |
author |
author2 |
Rossi, Danuza [UNESP] Silveira, Wagner da Silva [UNESP] Valentini, Sandro Roberto [UNESP] Zanelli, Cleslei Fernando [UNESP] |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Galvão, Fabio Carrilho [UNESP] Rossi, Danuza [UNESP] Silveira, Wagner da Silva [UNESP] Valentini, Sandro Roberto [UNESP] Zanelli, Cleslei Fernando [UNESP] |
dc.subject.por.fl_str_mv |
deoxyhypusine synthase deoxyhypusine synthase 1 deoxyhypusine synthase 1 1 fungal protein hypusine initiation factor 5A protein Asc1 protein kinase C sorbitol unclassified drug allele asc1 gene binding affinity cell growth cell wall cell wall integrity controlled study cytolysis deoxyhypusine synthase 1 gene eukaryotic translation initiation factor 5A gene gene gene function gene interaction gene overexpression genetic identification mutant nonhuman osmosis polysome protein depletion protein function protein kinase C gene protein modification protein protein interaction protein synthesis ribosome RNA translation sensitivity analysis translation regulation wild type yeast |
topic |
deoxyhypusine synthase deoxyhypusine synthase 1 deoxyhypusine synthase 1 1 fungal protein hypusine initiation factor 5A protein Asc1 protein kinase C sorbitol unclassified drug allele asc1 gene binding affinity cell growth cell wall cell wall integrity controlled study cytolysis deoxyhypusine synthase 1 gene eukaryotic translation initiation factor 5A gene gene gene function gene interaction gene overexpression genetic identification mutant nonhuman osmosis polysome protein depletion protein function protein kinase C gene protein modification protein protein interaction protein synthesis ribosome RNA translation sensitivity analysis translation regulation wild type yeast |
description |
The putative eukaryotic translation initiation factor 5A (eIF5A) is a highly conserved protein among archaea and eukaryotes that has recently been implicated in the elongation step of translation. eIF5A undergoes an essential and conserved posttranslational modification at a specific lysine to generate the residue hypusine. The enzymes deoxyhypusine synthase (Dys1) and deoxyhypusine hydroxylase (Lia1) catalyze this two-step modification process. Although several Saccharomyces cerevisiae eIF5A mutants have importantly contributed to the study of eIF5A function, no conditional mutant of Dys1 has been described so far. In this study, we generated and characterized the dys1-1 mutant, which showed a strong depletion of mutated Dys1 protein, resulting in more than 2-fold decrease in hypusine levels relative to the wild type. The dys1-1 mutant demonstrated a defect in total protein synthesis, a defect in polysome profile indicative of a translation elongation defect and a reduced association of eIF5A with polysomes. The growth phenotype of dys1-1 mutant is severe, growing only in the presence of 1 M sorbitol, an osmotic stabilizer. Although this phenotype is characteristic of Pkc1 cell wall integrity mutants, the sorbitol requirement from dys1-1 is not associated with cell lysis. We observed that the dys1-1 genetically interacts with the sole yeast protein kinase C (Pkc1) and Asc1, a component of the 40S ribosomal subunit. The dys1-1 mutant was synthetically lethal in combination with asc1Δ and overexpression of TIF51A (eIF5A) or DYS1 is toxic for an asc1Δ strain. Moreover, eIF5A is more associated with translating ribosomes in the absence of Asc1 in the cell. Finally, analysis of the sensitivity to cell wall-perturbing compounds revealed a more similar behavior of the dys1-1 and asc1Δ mutants in comparison with the pkc1Δ mutant. These data suggest a correlated role for eIF5A and Asc1 in coordinating the translational control of a subset of mRNAs associated with cell integrity. © 2013 Galvão et al. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-04-01 2014-05-27T11:28:48Z 2014-05-27T11:28:48Z |
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/10.1371/journal.pone.0060140 PLoS ONE, v. 8, n. 4, 2013. 1932-6203 http://hdl.handle.net/11449/74994 10.1371/journal.pone.0060140 WOS:000316930900050 2-s2.0-84875686470 2-s2.0-84875686470.pdf 5333250355049814 1525665408900195 0000-0001-7831-1149 |
url |
http://dx.doi.org/10.1371/journal.pone.0060140 http://hdl.handle.net/11449/74994 |
identifier_str_mv |
PLoS ONE, v. 8, n. 4, 2013. 1932-6203 10.1371/journal.pone.0060140 WOS:000316930900050 2-s2.0-84875686470 2-s2.0-84875686470.pdf 5333250355049814 1525665408900195 0000-0001-7831-1149 |
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 |
application/pdf |
dc.source.none.fl_str_mv |
Scopus 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_ |
1799965685205434368 |