Biophysical investigation of the iron in Aft1-1(up) and Gal-YAH1 Saccharomyces cerevisiae.
Biophysical investigation of the iron in Aft1-1(up) and Gal-YAH1 Saccharomyces cerevisiae.
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DOI：
10.1021/bi102015s
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发表时间：
2011-04-05
期刊：
影响因子：
2.9
通讯作者：
Lindahl PA
中科院分区：
文献类型：
--
作者：
Miao R;Holmes-Hampton GP;Lindahl PA
Aft 1 p是芽殖酵母Saccharomycescerevisiae中的主要铁调节因子。它间接感觉胞质铁状态，并通过激活或抑制铁调节子基因作出反应。Aft 1 -1up菌株中的Aft 1 p具有单个氨基酸突变，该突变导致其组成性激活铁调节子基因，而不管细胞Fe状态如何。这导致在低Fe和高Fe生长条件下升高的Fe吸收。铁氧还蛋白Yah 1 p参与Fe/S簇组装，Aft 1 p靶向铁调节子基因也在Yah 1 p缺失的细胞中上调。在这项研究中，穆斯堡尔谱，EPR和紫外-可见光谱被用来表征铁分布在Aft 1 -1up和Yah 1 p耗尽的细胞。在低铁培养基中生长的Aft 1 -1up细胞比WT细胞含有更多的铁。WT和Aft 1 -1up细胞中的基础水平的Fe位于线粒体中，主要以Fe/S簇和血红素中心的形式存在。Aft 1 -1up细胞中额外的Fe以HS Fe（III）形式存在。这些物种位于非线粒体位置，此处假设为空泡。在高铁培养基中生长的Aft 1 -1up细胞比WT细胞含有更多的铁。额外的铁是作为HS Fe（III）离子，可能储存在液泡中，和作为Fe（III）（磷酸盐）纳米粒子，位于线粒体中。Yah 1 p缺陷细胞也在线粒体中积累了纳米颗粒，但它们不含HS Fe（III）物质。结果解释了一个拟议的模型，涉及三个稳态调节系统，包括Aft 1系统，液泡铁调节系统和线粒体铁调节系统。
Aft1p is a major iron regulator in budding yeast Saccharomyces cerevisiae. It indirectly senses cytosolic Fe status and responds by activating or repressing iron regulon genes. Aft1p within the Aft1-1up strain has a single amino acid mutation which causes it to constitutively activate iron regulon genes regardless of cellular Fe status. This leads to elevated Fe uptake under both low and high Fe growth conditions. Ferredoxin Yah1p is involved in Fe/S cluster assembly, and Aft1p-targeted iron regulon genes are also up-regulated in Yah1p-depleted cells. In this study Mössbauer, EPR, and UV-vis spectroscopies were used to characterize the Fe distribution in Aft1-1up and Yah1p-depleted cells. Aft1-1up cells grown in low-Fe medium contained more Fe than do WT cells. A basal level of Fe in both WT and Aft1-1up cells was located in mitochondria, primarily in the form of Fe/S clusters and heme centers. The additional Fe in Aft1-1up cells was present as HS Fe(III) species. These species are in a non-mitochondrial location, assumed here to be vacuolar. Aft1-1up cells grown in high-Fe medium contained far more Fe than found in WT cells. The extra Fe was present as HS Fe(III) ions, probably stored in vacuoles, and as Fe(III) (phosphate) nanoparticles, located in mitochondria. Yah1p-deficent cells also accumulated nanoparticles in their mitochondria, but they did not contain HS Fe(III) species. Results are interpreted by a proposed model involving three homeostatic regulatory systems, including the Aft1 system, a vacuolar iron regulatory system and a mitochondrial Fe regulatory system.
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通讯作者：
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