This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Our project studies how the yeast Saccharomyces cerevisiae handles the essential metal ion magnesium (Mg2+). Mg2+ homeostasis is important because hundreds of cellular components require this ion in order to function properly. On the other hand, unregulated high intracellular Mg2+ concentrations ([Mg2+]IC) may lead to uncontrolled cell proliferation (1). The central coordinating role of free Mg2+ in growth regulation of normal cells gave rise to the hypothesis that neoplastic cells have lost their capacity to regulate the availability of Mg2+. Translating this situation to yeast, the lack of important Mg2+-homeostasis regulatory genes will cause a significantly higher rate of proliferation and subsequently an abnormally elevated final cell density (2). We are using functional genomics technology to identify genes involved in the uptake, distribution and storage of Mg2+ in the yeast cell as well as overall control of Mg2+ availability. Specifically, we are conducting genome-wide screenings at different extracellular [Mg2+] ([Mg2+]EC) since any single viable mutant with impaired Mg2+-homeostasis will exhibit a visible growth defect under low or high [Mg2+]EC. After identifying suitable mutants, we will first confirm: (a) the phenotype by conducting growth curves under different conditions, (b) the role of the deleted gene by reverting the phenotype. We will also determine the intracellular distribution of the metal ion, subcellular localization of the gene product, metal ion influx into the corresponding organelle and protein-protein interactions. Data gathered from these experimental should allow us to establish the biological function of these genes.
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