Proteomic Construction of Stress Regulated Networks

超出所提供的空间。所有生物都面临着一个普遍的挑战，那就是识别并试图抵消环境压力的有害影响。无论是多细胞生物还是单细胞生物，从细菌到哺乳动物，所有生物都进化出了应对这种分子攻击的机制。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）等因素的反应的基本机制在分子水平上是高度进化保守的。所有这些分子-细胞保护机制的一个主要特征是，某些细胞基因编码的蛋白质的转录被诱导，这些蛋白质具有应激保护特性。同时，编码缺乏保护性细胞/分子作用的蛋白质（和rna）的基因转录受到抑制。这些关键的转录反应都是通过一些独特的转录激活因子和抑制蛋白的作用来介导的。在这项研究中，我们建议使用最先进的多维串联质谱方法来鉴定和分子表征蛋白质-蛋白质调节网络，这些网络将环境与已知的关键保守转录因子连接起来，以驱动细胞RNA合成中基因水平的保护性变化。我们的研究将首先集中在遗传上易于处理和简单的真核生物贝克酵母（酿酒酵母）上。然而，在这个项目的后期，我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。网站性能 ======================================== 节结束 ===========================================

EXCEED THE SPACE PROVIDED. A universal challenge faced by all livingorganisms is to identify and then attept to counteract the deleterious effects of environmental stresses. Whether multicellular or unicellular, all organisms ranging from bacteria to mammals, 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 evolutionarily conserved at the molecularlevel. A cardinal feature of all of these molecular-cellularprotectivemechanisms is that transcriptionof certain cellulargenes encoding proteinsthat conferstress-protective properties is induced. Concomitantly,transcriptionof genes encoding proteins (and RNAs) that lack protective cellular/moleculareffects is repressed. These criticaltranscriptionalresponsesare all mediated throughthe actions of a handfulof distincttranscriptionalactivator and repressor proteins. In this studywe propose to use state of the art multidimensionaltandem mass spectrometry methods to identify and molecularly characterize the protein-proteinregulatory networksconnectingthe environmentwith key conserved transcriptionfactors knownto drive the gene-level protectivechanges in cellular RNA synthesis. Our studies will initiallyfocus uponthe geneticallytractable and simple eukaryote Baker'syeast (Saccharomyces cerevisiae). However, inthe latter phase of this project we will apply the knowledgegained in the unicellular yeast model system to human cells. PERFORMANCE SITE ========================================Section End===========================================