Molecular mechanisms of iron homeostasis in Saccharomyces cerevisiae

酿酒酵母铁稳态的分子机制

基本信息

  • 批准号:
    7486540
  • 负责人:
  • 金额:
    $ 2.79万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2008
  • 资助国家:
    美国
  • 起止时间:
    2008-04-01 至 2010-03-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Iron (Fe) deficiency is a nutritional disorder that affects nearly two billion people worldwide. The long-term goal of the proposed research is to understand how organisms cope with Fe-deficiency using the yeast Saccharomyces cerevisiae as a model system. The specific hypothesis is that the mRNA-binding proteins Cth1 and Cth2 cooperatively mediate the adaptation to Fe-limiting conditions in S. cerevisiae through promoting targeted mRNA decay. This hypothesis is based on the observations that 1) during Fe-deficiency, Cth2 promotes the down regulation of many mRNAs involved in metabolic pathways, and cth2 mutant cells grow poorly on medium lacking Fe; 2) deletion of the CTH1 gene on a cth2 strain further exacerbates the growth phenotype on low Fe conditions and cth1cth2 mutants fail to down regulate several of the mRNAs targeted for turnover during Fe-deficiency; and 3) concomitantly with the down regulation of many mRNAs, wild type cells show increased steady-state levels of glucose-regulated genes under low Fe, but not cth1cth2 cells under the same conditions. The specific aims are to: 1. Establish the contribution of Cth1 protein to the response to Fe limitation. The transcriptional activation, mRNA-binding and degradation-promoting activities of Cth1 in response to Fe-deficiency will be ascertained using traditional techniques such Chromatin ImmunoPrecipitation (ChIP), beta-galactosidase reporter assays, yeast three-hybrid assays and RNA blotting. Microarray analyses will be performed to identify specific targets of Cth1 and Cth2 in response to low Fe. 2. Establish the effects of Cth1 and Cth2 activity on glucose metabolism during Fe-deficiency. Preliminary data suggest that Fe-starved yeast express several glucose-repressed genes, but fail to do so in the absence of Cth1 and Cth2. Glucose utilization during Fe-deficiency will be monitored using enzymatic assays as well as examining phospho-Snfl and fructose-1, 6-bisphosphatase levels in wild type cells and in cth1cth2 mutants. In addition, non-mRNA binding alleles of Cth1 and Cth2 will be used to ascertain whether their decay-promoting activity is necessary for the secondary glucose starvation response. Iron (Fe) deficiency is the number one nutritional disorder affecting nearly two billion people worldwide. The goal of this research is to understand how eukaryotes adjust their metabolism to conditions of Fe-deficiency using the genetically tractable yeast Saccharomyces cerevisiae, which will contribute to a better understanding of how humans cope with cellular Fe-deficiency at the molecular level.
描述(由申请人提供):铁(Fe)缺乏症是一种营养失调,影响着全球近20亿人。该研究的长期目标是利用酿酒酵母作为模型系统来了解生物体如何应对缺铁。具体假设是,mRNA结合蛋白Cth1和Cth2通过促进靶向mRNA衰变,协同介导酿酒酵母对限铁条件的适应。这一假设基于以下观察结果:1)缺铁时,Cth2促进了许多参与代谢途径的mrna的下调,Cth2突变体细胞在缺铁环境下生长不良;2) CTH1基因的缺失进一步加剧了cth2菌株在低铁条件下的生长表型,并且在缺铁条件下,cth1cth2突变体不能下调一些靶向转换的mrna;3)随着许多mrna的下调,野生型细胞在低铁条件下葡萄糖调节基因的稳态水平增加,而在相同条件下,cth1cth2细胞则没有。具体目标是:

项目成果

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Sandra Viviana Vergara其他文献

Sandra Viviana Vergara的其他文献

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{{ truncateString('Sandra Viviana Vergara', 18)}}的其他基金

Molecular mechanisms of iron homeostasis in Saccharomyces cerevisiae
酿酒酵母铁稳态的分子机制
  • 批准号:
    7608664
  • 财政年份:
    2008
  • 资助金额:
    $ 2.79万
  • 项目类别:

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