title: Requirement of glutathione for Sod1 activation during lifespan extension.
abstract: It has been shown that the activation of cytosolic superoxide dismutase (Sod1) in Saccharomyces cerevisiae is only dependent on Ccs1, which is responsible for insertion of copper into the enzyme catalytic center, and that glutathione (GSH) is not necessary for this process. In this work, we addressed an important role of GSH in Sod1 activation by a Ccs1-dependent mechanism during oxidative stress and its role in yeast lifespan. Exponential cells of Saccharomyces cerevisiae, treated or not with 0.5 mM menadione for 1 h, were used for evaluation of the effect of a mild oxidative stress pre-treatment on chronological lifespan. The results showed that menadione induced a lifespan extension in the wild-type (WT) strain but this adaptive response was repressed in gsh1 and in sod1 strains. Interestingly, menadione treatment increased SOD1 and CCS1 gene expression in both WT and gsh1 strains. However, while these strains showed the same Sod1 activity before treatment, only the WT presented an increase of Sod1 activity after menadione exposure. Glutathionylation seems to be essential for Sod1 activation since no increase in activity was observed after menadione treatment in grx1 and grx2 null mutants. Our results suggest that GSH and glutathionylation are fundamental to protect Sod1 sulfhydryl residues under mild oxidative stress, enabling Sod1 activation and lifespan extension.
authors.affiliation: Departamento de Bioquímica, Instituto de Química, UFRJ, 21941-909, Rio de Janeiro, Brazil. sergiocantum@gmail.com
authors.forename: Sérgio C
authors.initials: SC
authors.lastname: Mannarino
authors.forename: Leonardo F
authors.initials: LF
authors.lastname: Vilela
authors.forename: Aline A
authors.initials: AA
authors.lastname: Brasil
authors.forename: Juliana N
authors.initials: JN
authors.lastname: Aranha
authors.forename: Pedro
authors.initials: P
authors.lastname: Moradas-Ferreira
authors.forename: Marcos D
authors.initials: MD
authors.lastname: Pereira
authors.forename: Vítor
authors.initials: V
authors.lastname: Costa
authors.forename: Elis C A
authors.initials: EC
authors.lastname: Eleutherio
mesh_terms: D002910:Chronology as Topic; D003300:Copper / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D054477:Glutaredoxins / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D018384:Oxidative Stress; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism*; D024483:Vitamin K 3 / Q000494:pharmacology*
chemical_list: C491960:CCS1 protein, S cerevisiae; D054477:Glutaredoxins; C530831:Grx1 protein, S cerevisiae; C527885:Grx2 protein, S cerevisiae; D018832:Molecular Chaperones; D029701:Saccharomyces cerevisiae Proteins; D024483:Vitamin K 3; D003300:Copper; D013482:Superoxide Dismutase; D005978:Glutathione