Sorting and Transport of Yeast Membrane Proteins

酵母膜蛋白的分选和运输

基本信息

批准号：
8899558
负责人：
Tom Hall Stevens
金额：
$ 35.25万
依托单位：
UNIVERSITY OF OREGON
依托单位国家：
美国
项目类别：
财政年份：
1987
资助国家：
美国
起止时间：
1987-04-01 至 2017-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8899558
关键词：
ATP Hydrolysis ATP phosphohydrolase Address Affect Albers-Schonberg disease Alleles Animal Model Animals Back Binding Biochemical Biological Assay Bone Diseases Brain Cells Collaborations Complementary DNA Complex Coupled Couples Cutis Laxa Defect Development Disease Drug Targeting Endoplasmic Reticulum Endosomes Eukaryota Eukaryotic Cell Foundations Genes Genetic Genetic Screening Goals Golgi Apparatus Grant Health Human Kidney Kidney Diseases Kinetics Knowledge Liver Medical center Membrane Membrane Protein Traffic Membrane Proteins Modeling Molecular Molecular Machines Muscle Mutation Neoplasm Metastasis Netherlands Organelles Orthologous Gene Pathway interactions Process Protein Isoforms Protein Subunits Proteins Proteolipids Proton-Translocating ATPases Protons Renal tubular acidosis Reporter Research Retrieval Role Saccharomyces cerevisiae Set protein Signal Transduction Sorting - Cell Movement System Translating Universities Vacuole Work Yeast Model System Yeasts base bone candidate identification clinical phenotype follow-up genetic analysis genome-wide genome-wide analysis human disease insight membrane assembly mutant plant fungi professor protein aminoacid sequence protein complex protein function protein protein interaction receptor tumor vacuolar H+-ATPase

项目摘要

DESCRIPTION (provided by applicant): This project will explore the mechanisms that govern the assembly, sorting and transport of the vacuolar- type proton-translocating ATPase (V-ATPase) in the simple model eukaryote, the yeast Saccharomyces cerevisiae. The V-ATPase is a molecular machine that couples the hydrolysis of ATP with the translocation of protons into the lumen of organelles, and the V-ATPase is highly conserved across fungi, plants and animals. Our genetic analysis in yeast has identified a group of genes in yeast encoding proteins that function in the endoplasmic reticulum (ER) in assembly of the membrane sector of the V-ATPase. These V-ATPase assembly factors will be characterized by genetic and biochemical approaches, to characterize the assembly pathway for the V-ATPase. Whereas it has been known for years that the V-ATPase is highly conserved, it has only very recently become clear that the V-ATPase assembly factors that we have identified in yeast are conserved in humans. Three specific aims are proposed. The first aim focuses on the important question of how each of the V- ATPase assembly factors functions in the highly orchestrated assembly of the 6-subunit integral membrane domain of the V-ATPase in the ER. This aim is also focused on investigating the mechanism of ER exit of the V-ATPase, and the function of the proteins identified as required for ER exit of this protein complex. The second aim centers on characterizing the human V-ATPase assembly factors in yeast. Our collaboration with Professor Dirk Lefeber has provided us with the cDNAs of four human V-ATPase assembly factors (hVma12, hVma21, hVma22 and Ac45), as well as unpublished information on mutant alleles of these genes that cause human disease. These proteins will be characterized for V-ATPase assembly function in the much simpler yeast model system. Finally, the third aim addresses the sorting and retention of the Golgi-endosomal form of the yeast V-ATPase that contains the second isoform of subunit "a", the Stv1 protein. We will use the knowledge of the Stv1 sorting/retention signal that we have recently identified to investigate and characterize the proteins involved in the sorting and retention of the Stv1-containing V-ATPase complex. The knowledge gained from these studies will provide new insights into the assembly, transport and sorting of this complex protein machine, and will provide insights into the mechanistic basis for the growing number of V-ATPase associated human diseases.

描述（由申请人提供）：该项目将探讨在简单模型真核生物，酵母菌糖酵母中，液泡型质子转移ATPase（V-ATPase）的组装，分类和运输的机制。 V-ATPase是一种分子机器，将ATP的水解与质子的易位到细胞器的腔内，并且V-ATPase在真菌，植物和动物之间高度保守。我们在酵母中的遗传分析已经确定了编码蛋白质的一组基因，该蛋白质在V-ATPase的膜扇区组装中在内质网（ER）中起作用。这些V-ATPase组装因子将以遗传和生化方法为特征，以表征V-ATPase的组装途径。多年来，人们已经知道V-ATPase是高度保守的，但最近才清楚地表明，我们在酵母中鉴定出的V-ATPase组装因子在人类中是保守的。提出了三个具体目标。第一个目的侧重于一个重要的问题，即每个V- ATPase组装因子如何在ER中V-ATPase的6-亚基积分膜结构域的高度策划组装中起作用。该目标还集中在研究V-ATPase的ER出口机理上，以及该蛋白质复合物ER出口所需的蛋白质的功能。第二个目的是表征酵母中人类V-ATPase组装因子。我们与Dirk Lefeber教授的合作为我们提供了四个人类V-ATPase组装因子（HVMA12，HVMA21，HVMA22和AC45）的CDNA，以及有关这些引起人类疾病的这些基因突变等位基因的未发表信息。这些蛋白质将在更简单的酵母模型系统中为V-ATPase组装功能进行表征。最后，第三个目的是针对酵母V-ATPase的高尔基 - 粘体形式的分类和保留，该酵母V-ATPase包含亚基“ A”，STV1蛋白的第二个同工型。我们将使用最近确定的STV1排序/保留信号的知识来研究和表征涉及含STV1的V-ATPase复合物的分类和保留的蛋白质。从这些研究中获得的知识将为这款复杂蛋白质机的组装，运输和分类提供新的见解，并将为越来越多的V-ATPase相关人类疾病提供洞察力。