Regulation of ESCRT-III Activity in Yeast

酵母中 ESRT-III 活性的调节

• 批准号: 9276361
• 负责人: CHARLES G ODORIZZI
• 金额: $ 6.39万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9276361
• 关键词: Binding; Biochemical; Biogenesis; Biological Assay; Biological Models; Communicable Diseases; Complex; Data; Deubiquitination; Disease; Electron Microscopy; Electron Transport Complex III; Electrons; Endosomes; Functional disorder; Goals; HIV-1; Health; Hereditary Disease; Homeostasis; Human; Hydrolase; In Vitro; Infection; Integral Membrane Protein; Knowledge; Link; Lysosomes; Mediating; Membrane; Membrane Protein Traffic; Methodology; Microscopic; Molecular; Multivesicular Body; Nerve Degeneration; Neurodegenerative Disorders; Outcome; Pathway interactions; Phosphatidylinositols; Physiological; Prevention; Process; Protein Biochemistry; Proteins; Publishing; Reaction; Regulation; Research; Retroviridae; Role; Saccharomyces cerevisiae; Saccharomycetales; Site; Sorting - Cell Movement; Structure; Testing; Therapeutic; Transmembrane Domain; Ubiquitin; Vesicle; Work; Yeasts; base; combat; human disease; in vivo; innovation; insight; light microscopy; man; mutant; pathogen; phosphatidylinositol 3-phosphate; trafficking; yeast protein

DESCRIPTION (provided by applicant): The endosomal sorting complexes required for transport (ESCRTs) comprise a cellular machinery conserved from yeast to man. ESCRTs function in the biogenesis of late endosomal multivesicular bodies (MVBs) by executing the formation of intralumenal vesicles (ILVs), which bud directly from the endosomal membrane toward the lumen of the compartment. ILVs perform a critical role in maintaining cellular homeostasis by carrying unneeded, damaged, or dangerous transmembrane proteins to be degraded in the hydrolytic interior of the lysosome upon endolysosomal fusion. Dysfunction of the ESCRT-III complex has been linked to neurodegenerative disease, and its exploitation is essential for the spread of HIV-1 and other retroviruses as well as certain bacterial pathogens. In vitro studies have shown that ESCRT-III catalyzes membrane scission in the ILV budding pathway at MVBs. The objective of this research is to determine the regulatory mechanisms that control ESCRT-III activity in vivo, using the budding yeast Saccharomyces cerevisiae as a model system. The central hypothesis is that ESCRT-III activity is regulated in vivo by proteins that control the assembly and disassembly of this complex. This research will study how the structure and function of ESCRT-III is impacted in yeast by proteins that regulate assembly and disassembly of the complex. The methodology to be used includes electron microscopy, light microscopy, protein biochemistry, and functional assays. The rationale for this research is that the regulation of ESCRT-III must be defined if therapeutic strategies are to be developed that will treat ESCRT-III dysfunction or combat exploitations of ESCRT-III by human pathogens. Using yeast as a model system to understand this regulation is directly relevant to human health because the mechanism of ESCRT-III function is highly conserved. With respect to expected outcomes, it is anticipated that completion of this research will reveal the core, broadly conserved mechanisms that regulate ESCRT-III activity. Such results are expected to have an important positive impact because they will yield fundamental insight into a poorly understood regulatory step in the endocytic pathway, reveal potential targets for the prevention and treatment of diseases linked to ESCRT-III, and inspire new and innovative approaches to understand the mechanisms of ESCRT-III function.

描述（由申请人提供）：转运所需的内体分选复合物（ESCRT）包含从酵母到人保守的细胞机制。ESCRT通过形成腔内小泡（ILV）在晚期内体多泡体（MVB）的生物发生中发挥作用，ILV直接从内体膜向腔室的内腔出芽。ILV通过携带不需要的、受损的或危险的跨膜蛋白在溶酶体内融合时在溶酶体的水解内部降解，在维持细胞稳态中发挥关键作用。ESCRT-III复合物的功能障碍与神经退行性疾病有关，其开发对于HIV-1和其他逆转录病毒以及某些细菌病原体的传播至关重要。体外研究表明，ESCRT-III在MVB的ILV出芽途径中催化膜断裂。本研究的目的是确定的监管机制，控制ESCRT-III的活性在体内，使用芽殖酵母酿酒酵母作为模型系统。核心假设是ESCRT-III活性在体内由控制该复合物组装和分解的蛋白质调节。这项研究将研究ESCRT-III的结构和功能如何在酵母中受到调节复合物组装和拆卸的蛋白质的影响。使用的方法包括电子显微镜、光学显微镜、蛋白质生物化学和功能测定。这项研究的基本原理是，如果要开发治疗ESCRT-III功能障碍或对抗人类病原体对ESCRT-III的利用的治疗策略，则必须定义ESCRT-III的调节。使用酵母作为模型系统来理解这种调节与人类健康直接相关，因为ESCRT-III功能的机制是高度保守的。关于预期的结果，预计这项研究的完成将揭示调节ESCRT-III活性的核心、广泛保守的机制。这些结果预计将产生重要的积极影响，因为它们将产生对内吞途径中一个知之甚少的调控步骤的基本见解，揭示预防和治疗与ESCRT-III相关疾病的潜在目标，并激发新的创新方法来了解ESCRT-III功能的机制。