The role of the Saccharomyces cerevisiae CCC1 gene in the homeostasis of manganese ions

Abstract

We previously reported that oxidative damage in yeast lacking copper/zinc superoxide dismutase (SOD1) can be alleviated through mutations in PMR1, encoding a calcium P-type ATPase homologue that also functions in manganese homeostasis. In an attempt to further understand the relationship between manganese ions, PMR1 and SOD1, we conducted a search for manganese homeostasis genes that interact with PMR1. A genomic library was screened for genes that, when overexpressed, suppress the manganese hypersensitivity associated with pmr1 mutations. A single clone was isolated that reduced manganese toxicity in both the pmr1 mutant and PMR1 wild-type yeast. This gene was identified as CCC1, previously shown to function in calcium metabolism. Our studies indicate that, like PMR1, CCC1 functions in the homeostasis of both calcium and manganese ions. The Ccc1p polypeptide was found to localize to a Golgi-like organelle in yeast cells. Ccc1 p co-fractionated with a Golgi marker in subcellular fractionation studies and, with immunofluorescence microscopy, Ccc1 p exhibited a punctate pattern of staining typical of yeast Golgi. Our studies suggest that Ccc1 p may act to sequester manganese ions in this organelle and limit the intracellular availability of the metal. First, overexpression of CCC1 reduced manganese cytotoxicity without lowering total accumulation of the metal. Second, overexpression of CCC1 appeared to limit the intracellular availability of the manganese ions needed to support aerobic growth of SOD1 mutants. We provide a model in which Ccc1p and Pmr1p work together to control the intracellular partitioning of manganese ions.