ANALYSIS OF GOLGI MEMBRANE PROTEIN LOCALIZATION IN YEAST

酵母高尔基体膜蛋白定位分析

• 批准号: 6525902
• 负责人: Steven F Nothwehr
• 金额: $ 18.18万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6525902
• 关键词: Golgi apparatus; Saccharomyces cerevisiae; actins; alkaline phosphatase; aminopeptidase; enzyme activity; fungal proteins; gene mutation; intracellular transport; laboratory rabbit; membrane proteins; phosphoprotein phosphatase; phosphoproteins; protein structure function; protein transport

The sorting and localization of proteins in the secretory and endosomal pathways will be investigated, using the model system of localization of proteins to the TGN in S. cerevisiae. The PI's working model proposes two independent mechanisms that both act to localize proteins to the TGN: selective retrieval from the prevacuolar compartment and a reduced rate of transport out to the TGN. Preliminary data suggest that retrieval involves the interaction of cargo proteins with Vps35p, a prevacuolar peripheral membrane protein, and this hypothesis will be tested in a genetic suppressor approach, as well as in biochemical binding assays. Slow exit from the TGN is proposed to involve the phosphorylation of cytosolic domains of resident TGN proteins, and this will be investigated using biochemical approaches to identify the specific sites of phosphorylation followed by specific site mutagenesis to alter these phosphorylation sites. Finally, several genes have been identified in genetic screens as defective in Golgi retention of proteins, and four of these will be investigated in further detail: grd20, grd21, grd22 and grd23.

蛋白质中蛋白质的分类和定位 使用定位模型系统，将研究内体途径 在酿酒酵母中蛋白质的蛋白质。 PI的工作模型提出了两个 两者都作用于将蛋白质定位到TGN的独立机制：选择性 从prevacuolol室检索和降低的运输速率 到TGN。初步数据表明，检索涉及相互作用 带有VPS35P的货物蛋白，一种prevacuolar外围膜蛋白，这是 假设将以遗传抑制方法以及在 生化结合测定。提议从TGN退出缓慢的退出 常驻TGN蛋白的胞质结构域的磷酸化，这将是 使用生化方法进行了研究以识别 磷酸化，然后进行特定的位点诱变，以改变这些 磷酸化位点。最后，在遗传中鉴定了几个基因 蛋白质高尔基保留率有缺陷的筛选，其中四个将是 进一步研究：GRD20，GRD21，GRD22和GRD23。