Copper transport inside the cells is important since this transition metal catalyzes many reactions required for proper cellular functions. This mechanism in Saccharomyces cerevisiae is controlled by the copper transport protein (Ctr1p), a membrane protein that regulates this high affinity copper transport machinery by assisting and bringing copper to the intracellular compartment. In addition, other three major proteins (Cox17p, Atx1p, and Ccs1p) serve as chaperones routing this metal to specific organelles. In order to elucidate the copper transfer mechanism in yeast, wild type (wt), ctr1Δ and atx1Δ mutant cells were grown under low and rich copper conditions. These conditions were achieved by treating cells with the chelating agent bathocuproinedisulfonate (BCS) and copper (II) sulfate (CuSO4), respectively. Cell growth sensitivity assays revealed that both ctr1Δ and wt cells grew normal under copper deficient conditions. However, atx1Δ cells, which lack targeting of copper to the secretory compartment, grew significantly faster at higher copper-deficient conditions. This data suggest that copper deprivation on atx1Δ cells activates the high affinity copper transport machinery, increasing the copper uptake, hence the cell growth increase in these mutant cells. On the other hand, our results showed that CuSO4 strongly affected the growth in wt and ctr1Δ cells. However, this decrease in growth was strongly observed in wt compared ctr1Δ mutant cells. These results suggest that since copper is not being effectively recruited in ctr1Δ mutant cells, deletion of the CTR1 gene provide higher resistance in rich copper conditions. Elucidation of the copper transport machinery in Saccharomyces cerevisiae will allow us to identify target genes involved in the response to copper imbalance associated with health conditions in humans. This research is supported by an Institutional Development Award (IDeA) from the NIGMS of the NIH (8P20GM103475-12) and the PRLSAMP Program from the (NSF).

Effect of copper imbalance in cell viability of the budding yeast Saccharomyces cerevisiae

251st National Meeting (2016)

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