Mutations of Cys and Ser residues in the alpha5-subunit of the 20S proteasome from Saccharomyces cerevisiae affects gating and chronological lifespan

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dc.contributorLab. Bioquímicapt_BR
dc.contributor.authorLeme, Janaina de Moraes Macielpt_BR
dc.contributor.authorOhara, Erinapt_BR
dc.contributor.authorSantiago, Verônica Feijolipt_BR
dc.contributor.authorBarros, Mario H.pt_BR
dc.contributor.authorNetto, Luis E. S.pt_BR
dc.contributor.authorPimenta, Daniel Carvalhopt_BR
dc.contributor.authorMariano, Douglas Oscar Ceolinpt_BR
dc.contributor.authorOliveira, Cristiano L. P.pt_BR
dc.contributor.authorBicev, Renata N.pt_BR
dc.contributor.authorBarreto-Chaves, Maria L. M.pt_BR
dc.contributor.authorLino, Caroline A.pt_BR
dc.contributor.authorDemasi, Marilenept_BR
dc.identifier.citationLeme JMM, Ohara E, Santiago VF, Barros MH., Netto LE.S., Pimenta DC, et al. Mutations of Cys and Ser residues in the alpha5-subunit of the 20S proteasome from Saccharomyces cerevisiae affects gating and chronological lifespan. Arch Biochem Biophys. 2019 May;666:63-72. doi:10.1016/
dc.description.abstractIn addition to autophagy, proteasomes are critical for regulating intracellular protein levels and removing misfolded proteins. The 20S proteasome (20SPT), the central catalytic unit, is sometimes flanked by regulatory units at one or both ends. Additionally, proteosomal activation has been associated with increased lifespan in many organisms. Our group previously reported that the gating (open/closed) of the free 20S proteasome is redox controlled, and that S-glutathionylation of two Cys residues (Cys76 and Cys221) in the alpha5 subunit promotes gate opening. The present study constructed site-directed mutants of these Cys residues, and evaluated the effects these mutations have on proteosome gate opening and yeast cell survival. Notably, the double mutation of both Cys residues (Cys76 and Cys221) rendered the cells nonviable, whereas the lifespan of the yeast carrying the single mutations (alpha5-C76S or alpha5-C221S) was attenuated when compared to the wild type counterpart. Furthermore, it was found that alpha5-C76S or alpha5-C221S 20SPT were more likely to be found with the gate in a closed conformation. In contrast, a random alpha5-subunit double mutation (S35P/C221S) promoted gate opening, increased chronological lifespan and provided resistance to oxidative stress. The 20SPT core particle purified from the long-lived strain degraded model proteins (e.g., a-synuclein) more efficiently than preparations obtained from the wild-type counterpart, and also displayed an increased chymotrypsin-like activity. Mass spectrometric analyses of the C76S, C221S, S35P/C221S, S35P and S35P/C76S mutants provided evidence that the highly conserved Cys76 residue of the alpha5-subunit is the key determinant for gate opening and cellular survival. The present study reveals a sophisticated regulatory mechanism that controls gate opening, which appears to be based on the interactions among multiple residues within the a5-subunit, and consequently impacts the lifespan of yeast.pt_BR
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)pt_BR
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)pt_BR
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)pt_BR
dc.format.extentp. 63-72pt_BR
dc.relation.ispartofArchives of Biochemistry and Biophysicspt_BR
dc.titleMutations of Cys and Ser residues in the alpha5-subunit of the 20S proteasome from Saccharomyces cerevisiae affects gating and chronological lifespanpt_BR
dc.contributor.externalUniversidade de São Paulo (USP)¦¦Brasilpt_BR
dc.subject.keywordChronological lifespanpt_BR
dc.subject.keywordPost-translational modificationpt_BR
dc.subject.keywordRedox regulationpt_BR
dc.relation.ispartofabbreviatedArch Biochem Biophyspt_BR
dc.identifier.citationabntv. 666, p. 63-72, maio 2019pt_BR
dc.identifier.citationvancouver2019 May;666:63-72pt_BR
dc.contributor.butantanLeme, Janaina de Moraes Maciel|:Aluno|:Lab. Bioquímica|:PrimeiroAutorpt_BR
dc.contributor.butantanPimenta, Daniel Carvalho|:Pesquisador:Docente Permanente PPGTOX|:Lab. Bioquímica|:pt_BR
dc.contributor.butantanOhara, Erina|:Aluno|:Lab. Bioquímica|:pt_BR
dc.contributor.butantanSantiago, Verônica Feijoli|:Aluno|:Lab. Bioquímica|:pt_BR
dc.contributor.butantanMariano, Douglas Oscar Ceolin|:Aluno|:Lab. Bioquímica|:pt_BR
dc.contributor.butantanDemasi, Marilene|:Pesquisador|:Lab. Bioquímica|:Autor de correspondênciapt_BR
dc.sponsorship.butantanConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)¦¦573530/2008-4pt_BR
dc.sponsorship.butantanCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)¦¦pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2014/21058-0pt_BR
dc.sponsorship.butantanFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)¦¦2013/07937-8pt_BR
dc.subject.researchlineToxinologia estruturalpt_BR
item.fulltextCom Texto completo-
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