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this experiment is based on a large-scale analysis of the yeast deletion mutant collection, using metal-specific viability assays. the extensive resources available for studying the yeast deletion collection were used to examine the extent to which genes that are involved in the cellular response to cadmium and nickel overlap. the results demonstrate the feasibility of a robust and high-throughput method for the characterization of cellular responses to metal toxicity. the information gleaned from this screen may be used to elucidate the mechanisms of metal tolerance and the effects of metal toxicity on human health. the genetic loci identified in this screen may also be used to identify components of the metal response that can be manipulated to provide protection from metal-induced toxicity, with particular relevance to the development of metal-specific drugs.
contrary to the general assumption that zinc uptake occurs in the cytoplasm, we have found that msc1p plays an important role in the metal ion accumulation process. msc1p, an important regulator of gene expression and a zinc-binding protein [25], is the sole zinc transporter in yeast. to determine the precise site of zinc entry into the cell, we monitored zinc accumulation in various mutants of zinc homeostasis. surprisingly, we found that the spt3 mutation, which is known to affect vacuolar sequestration of zinc [25], did not cause any decrease in zinc accumulation. this suggests that zinc is not transported into the vacuole, but is bound to a cytoplasmic zinc-binding protein. we also found that nrm1, the metalloregulator of [25], is not essential for zinc uptake. moreover, the nrm1 mutant showed similar zinc accumulation patterns to wild type, suggesting that the observed phenotype was not due to a lack of response to zinc homeostasis. to confirm this, we examined the effect of zinc on cell growth. under both zinc-replete and zinc-depleted conditions, zinc-deficient mutants (zrc1, zrc2, zrc4 and zrc5) exhibited growth defects. we also found that the zinc uptake machinery (msc1p, zrc1p, zrc2p, zrc4p and zrc5p) was constitutively expressed under both zinc-replete and zinc-depleted conditions. thus, the zinc-deficiency phenotypes were likely due to zinc transport defects, as opposed to an effect of zinc on the expression of the transporter.
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