Effect of the membrane protein Alr1 in cadmium absorption by Saccharomyces cerevisiae
Mathias, C.J.; Gomes, D.S.; Panek, A.D.; Henriques, J.A.P.; Eleutherio, E.C.A.
Departamento de Bioquímica, Instituto de Química, UFRJ
Besides having a large potential to remove heavy metals from the environment, the yeast Saccharomyces cerevisiae has been used as an eukaryotic cell model to investigate mechanisms that control metal sensibility. Previous results showed that yeast cells have a large cadmium absorption capacity of the medium. It was also observed that metal absorption depends on metabolism, because dead cells do not absorb cadmium. According to literature, the membrane protein Alr1 seems to be involved with Cd2+ efflux to environment. In this work, we investigate cadmium absorption ability of genetically modified strains that overexpress ALR1 by constructing strains with ALR1 under control of the GAL promoter. Cells were grown in minimal media containing glucose or galactose as carbon sources, collected in exponential phase, centrifuged and ressuspended in phosphate buffer (50 mM pH 6.0) supplied with 40 mM CdSO4. The capacity of cells of absorbing Cd2+ was monitored by atomic absorption spectroscopy. According to our results, when cells were grown overexpressing ALR1, they absorbed almost 2-fold less Cd2+ and showed increased tolerance, bringing support to the hypothesis that Alr1 is involved with Cd2+ efflux.
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