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J. Biol. Chem., Vol. 277, Issue 48, 45751-45758, November 29, 2002

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Intracellular Glucose 1-Phosphate and Glucose 6-Phosphate Levels Modulate Ca²⁺ Homeostasis in Saccharomyces cerevisiae^*

David P. Aiello, Lianwu Fu, Attila Miseta^§, and David M. Bedwell^¶

From the  Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294-2170 and the ^§ Department of Clinical Chemistry, University Medical School, Peçs 7624, Hungary

The enzyme phosphoglucomutase plays a key role in cellular metabolism by virtue of its ability to interconvert Glc-1-P and Glc-6-P. It was recently shown that a yeast strain lacking the major isoform of phosphoglucomutase (pgm2) accumulates a high level of Glc-1-P and exhibits several phenotypes related to altered Ca²⁺ homeostasis when D-galactose is utilized as the carbon source (Fu, L., Miseta, A., Hunton, D., Marchase, R. B., and Bedwell, D. M. (2000) J. Biol. Chem. 275, 5431-5440). These phenotypes include increased Ca²⁺ uptake and accumulation and sensitivity to high environmental Ca²⁺ levels. In the present study, we overproduced the enzyme UDP-Glc pyrophosphorylase to test whether the overproduction of a downstream metabolite produced from Glc-1-P can also mediate changes in Ca²⁺ homeostasis. We found that overproduction of UDP-Glc did not cause any alterations in Ca²⁺ uptake or accumulation. We also examined whether Glc-6-P can influence cellular Ca²⁺ homeostasis. A yeast strain lacking the -subunit of phosphofructokinase (pfk2) accumulates a high level of Glc-6-P (Huang, D., Wilson, W. A., and Roach, P. J. (1997) J. Biol. Chem. 272, 22495-22501). We found that this increase in Glc-6-P led to a 1.5-2-fold increase in total cellular Ca²⁺. We also found that the pgm2/pfk2 strain, which accumulated high levels of both Glc-6-P and Glc-1-P, no longer exhibited the Ca²⁺-related phenotypes associated with high Glc-1-P levels in the pgm2 mutant. These results provide strong evidence that cellular Ca²⁺ homeostasis is coupled to the relative levels of Glc-6-P and Glc-1-P in yeast.

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