The role of the yeast ATP-binding cassette Ycf1p in glutathione and cadmium ion homeostasis during respiratory metabolism.

Cadmium (Cd2+) is a toxic environmental contaminant for biological systems, which can form complexes with reduced glutathione (GSH), and thus alter the intracellular redox state. In Saccharomyces (S.) cerevisiae, bis(glutathionato)cadmium (Cd-[GS]2) complexes can be removed from the cytosol and transported into the vacuole by a glutathione-conjugated pump, Ycf1p. In this study, we investigated the role of Ycf1p in Cd2+ detoxification during respiratory metabolism of S. cerevisiae, and the correlation of Ycf1p with GSH intracellular homeostasis. The results showed that in respiratory condition the mutant ycf1Delta is more tolerant to Cd2+ and to the oxidants t-BOOH and H2O2 than wild-type strain. This tolerance is probably related to the high content of GSH present in ycf1Delta mutant. The expression of YCF1 promoter in the wild-type strain is naturally down-regulated after the transition from fermentative to respiratory metabolism (diauxic shift), and its induction in response to Cd2+ is dependent on GSH availability. Our data suggest that Ycf1p is involved in the maintenance of intracellular GSH homeostasis and it can interfere with the oxidative tolerance of yeast. Moreover, the detoxification of Cd2+ is dependent on GSH availability and on cellular metabolic status.

The Pmr1 protein, the major yeast Ca2+-ATPase in the Golgi, regulates intracellular levels of the cadmium ion.

Cadmium is a nonessential, highly toxic heavy metal that shows ionic properties similar to calcium. These ionic similarities imply that the cadmium ion, Cd2+, is a calcium ion, Ca2+, receptor-agonist, affecting the same biochemical pathways involved in Ca2+ homeostasis. In the yeast Saccharomyces cerevisiae, the PMC1 and PMR1 genes encode vacuolar and Golgi Ca2+-ATPases, respectively. The PMR1 protein product Pmr1p is involved in both Ca2+ and Mn2+ homeostasis. This study investigated the importance of Pmc1p and Pmr1p for Cd2+ cellular detoxification. Using the standard techniques of yeast molecular research and a multielemental procedure named particle-induced X-ray emission, Pmr1p was identified as a protein that directly participates in the detoxification of Cd2+, possibly through the secretory pathway. The results allow us to posit a model of Cd2+ detoxification where Pmr1p has a central role in cell survival in a Cd2+-rich environment.