Regulation of ESCRT-III Activity in Yeast

酵母中 ESRT-III 活性的调节

• 批准号: 8746988
• 负责人: CHARLES G ODORIZZI
• 金额: $ 30.61万
• 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/8746988
• 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; public health relevance; 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）：传输（ESCRTS）所需的内体分类复合物包括从酵母到人保守的蜂窝机械。通过执行内体囊泡（ILV）形成的内体多囊体（MVB）生物发生的ESCRT在生物发生中的功能，这些囊泡（ILV）直接从内体膜向腔室的腔内直接芽。 ILV通过携带不需要的，受损或危险的跨膜蛋白来维持细胞稳态，在内溶液融合后的水解内部降解。 ESCRT-III复合物的功能障碍与神经退行性疾病有关，其剥削对于HIV-1和其他逆转录病毒以及某些细菌病原体的扩散至关重要。体外研究表明，ESCRT-III催化MVBS的ILV萌芽途径中的膜分裂。这项研究的目的是确定使用酿酒酵母作为模型系统的芽酵母糖果疗法来确定体内控制ESCRT-III活性的调节机制。中心假设是通过控制该复合物的组装和拆卸的蛋白质在体内调节ESCRT-III活性。这项研究将研究如何通过调节复合物组装和拆卸的蛋白质影响ESCRT-III的结构和功能。要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分析。这项研究的基本原理是，如果要开发将处理ESCRT-III功能障碍或通过人类病原体对ESCRT-III进行ESCRT-III功能障碍或作战剥削，则必须定义ESCRT-III的调节。使用酵母作为模型系统来了解该调节与人类健康直接相关，因为ESCRT-III功能的机制高度保守。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。预计此类结果将产生重要的积极影响，因为它们将对内吞途径中理解的调节步骤的理解不足，揭示了预防与ESCRT-III相关的疾病的潜在靶标，并激发新的和创新的方法，以了解ESCRT-III功能的机制。