Simm Claudia, Lahner Brett, Salt David, LeFurgey Ann, Ingram Peter, Yandell Brian, Eide David J
Department of Nutritional Sciences, 1415 Linden Drive, University of Wisconsin-Madison, Madison, WI 53706, USA.
Eukaryot Cell. 2007 Jul;6(7):1166-77. doi: 10.1128/EC.00077-07. Epub 2007 May 25.
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 10(8) 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.
先前对酿酒酵母的研究表明,液泡是细胞中储存锌的主要场所。然而,这些研究并未涉及液泡中储存锌的绝对水平,也未检测细胞其他区室中储存锌的丰度。在本报告中,我们描述了一种利用细胞器分级分离法和电子探针X射线微分析法对细胞内锌分布进行的分析。通过这些方法,我们确定液泡中的锌水平会随锌状态而变化,并且可以升至近100 mM锌(即每个细胞有7×10⁸个液泡锌原子)。此外,在锌饥饿条件下,这种锌可以有效地被动员起来,以满足多达八代子代细胞的需求。虽然Zrc1和Cot1锌转运蛋白在稳态生长条件下对锌进入液泡至关重要,但在“锌冲击”期间,当锌受限的细胞重新获得锌供应时,其他转运蛋白有助于介导锌进入液泡。此外,我们发现细胞的其他区室不会提供大量的锌储存。特别是,线粒体中的锌积累量很低,并且其稳态调节独立于液泡锌储存。最后,我们观察到锌状态与细胞中积累的镁和磷水平之间存在很强的相关性。我们的研究结果表明锌是细胞储存这些其他重要营养素能力的主要决定因素。