Proteomic Construction of Stress Regulated Networks

描述(由申请人提供) 所有活着的有机体面临的一个普遍挑战是识别并试图抵消环境压力的有害影响。无论是多细胞的还是单细胞的，所有的生物都进化出了应对这种分子侮辱的机制。事实上，对电离辐射、氧化应激、诱变和有毒小分子、营养剥夺和极端温度(热休克)等物质的基本反应机制在分子水平上高度保守，在进化上也是如此。所有这些分子-细胞保护机制的一个基本特征是诱导某些细胞基因的转录，这些基因编码赋予压力保护特性的蛋白质。与此同时，编码缺乏细胞/分子保护作用的蛋白质(和RNA)的基因转录受到抑制。这些关键的转录反应都是通过少数不同的转录激活蛋白和抑制蛋白的作用来调节的。在这项研究中，研究人员建议使用最先进的多维串联质谱学方法来鉴定和表征蛋白质-蛋白质调控网络，该网络将环境与已知驱动细胞RNA合成中基因水平保护性变化的关键保守转录因子联系起来。这些研究最初将集中在遗传上易驯化和简单的真核生物面包酵母(酿酒酵母)。然而，在这个项目的后期，他们将把在单细胞酵母模型系统中获得的知识应用到人类细胞中。

DESCRIPTION (provided by the applicant) A universal challenge faced by all living organisms is to identify and then attempt to counteract the deleterious effects of environmental stresses. Whether multicellular or unicellular, all organisms have evolved mechanisms to deal with such molecular insults. Indeed the fundamental mechanisms for responding to such agents as ionizing radiation, oxidative stress, mutagenic and toxic small molecules, nutrient deprivation and temperature extremes (heat shock) have been highly and evolutionarily conserved at the molecular level. A cardinal feature of all of these molecular-cellular protective mechanisms is the induction of the transcription of certain cellular genes encoding proteins that confer stress-protective properties. Concomitantly, transcription of genes encoding proteins (and RNAs) that lack protective cellular/molecular effects is repressed. These critical transcriptional responses are all mediated through the actions of a handful of distinct transcriptional activator and repressor proteins. In this study, the investigators propose to use state-of-the-art multidimensional tandem mass spectrometry methods to identify and characterize the protein-protein regulatory networks connecting the environment with key conserved transcription factors known to drive the gene-level protective changes in cellular RNA synthesis. The studies will initially focus on the genetically tractable and simple eukaryote Baker's yeast (Saccharomyces cerevisiae). However, in the latter phase of this project, they will apply the knowledge gained in the unicellular yeast model system to human cells.