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Eukaryotic Cell, July 2007, p. 1166-1177, Vol. 6, No. 7
1535-9778/07/$08.00+0     doi:10.1128/EC.00077-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Saccharomyces cerevisiae Vacuole in Zinc Storage and Intracellular Zinc Distribution ^,

Claudia Simm,^1, Brett Lahner,² David Salt,² Ann LeFurgey,³ Peter Ingram,⁴ Brian Yandell,⁵ and David J. Eide^1*

Department of Nutritional Sciences, University of Wisconsin—Madison, Madison, Wisconsin 53706,¹ Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, Indiana 47907-2010,² Department of Cell Biology, Duke University Medical Center and Veterans Affairs Medical Center, Durham, North Carolina 27705,³ Department of Pathology, Duke University Medical Center, Durham, North Carolina 27710,⁴ Departments of Statistics and Horticulture, University of Wisconsin—Madison, Madison, Wisconsin 53706⁵

Received 13 March 2007/ Accepted 9 May 2007

Previous studies of the yeast Saccharomyces cerevisiae indicated that the vacuole is a major site of zinc storage in the cell. However, these studies did not address the absolute level of zinc that was stored in the vacuole nor did they examine the abundances of stored zinc in other compartments of the cell. In this report, we describe an analysis of the cellular distribution of zinc by use of both an organellar fractionation method and an electron probe X-ray microanalysis. With these methods, we determined that zinc levels in the vacuole vary with zinc status and can rise to almost 100 mM zinc (i.e., 7 x 10⁸ atoms of vacuolar zinc per cell). Moreover, this zinc can be mobilized effectively to supply the needs of as many as eight generations of progeny cells under zinc starvation conditions. While the Zrc1 and Cot1 zinc transporters are essential for zinc uptake into the vacuole under steady-state growth conditions, additional transporters help mediate zinc uptake into the vacuole during "zinc shock," when zinc-limited cells are resupplied with zinc. In addition, we found that other compartments of the cell do not provide significant stores of zinc. In particular, zinc accumulation in mitochondria is low and is homeostatically regulated independently of vacuolar zinc storage. Finally, we observed a strong correlation between zinc status and the levels of magnesium and phosphorus accumulated in cells. Our results implicate zinc as a major determinant of the ability of the cell to store these other important nutrients.

Published ahead of print on 25 May 2007.

Supplemental material for this article may be found at http://ec.asm.org/.

Present address: Department of Chemistry, Northwestern University, Evanston, IL 60208-3113.