Proteomic Construction of Stress Regulated Networks

描述（由申请人提供） 所有生物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，所有的生物体都进化出了应对这种分子损伤的机制。 事实上，对电离辐射、氧化应激、致突变和有毒小分子、营养缺乏和极端温度（热休克）等物质作出反应的基本机制在分子水平上高度保守，而且在进化上也是如此。 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，这些基因编码赋予应激保护特性的蛋白质。 伴随地，编码缺乏保护性细胞/分子作用的蛋白质（和RNA）的基因的转录被抑制。 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介导的。 在这项研究中，研究人员建议使用最先进的多维串联质谱方法来识别和表征蛋白质-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连接起来。 这些研究最初将集中在遗传上易于处理和简单的真核生物面包酵母（酿酒酵母）。 然而，在该项目的后期，他们将把单细胞酵母模型系统中获得的知识应用于人类细胞。

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.