Role for Protein Phosphorylation in Yeast Vacuolar Protein Sorting

蛋白质磷酸化在酵母液泡蛋白质分选中的作用

• 批准号: 9017678
• 负责人: Scott Emr
• 金额: $ 2.79万
• 依托单位: California Institute of Technology
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-07-01 至 1991-10-01
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9017678&HistoricalAwards=false
• 关键词: Role; Protein; Phosphorylation; Yeast; Vacuolar

The proposed studies focus on one of the yeast vacuolar protein sorting (VPS) genes, VPS15. The predicted product of the VPS15 gene shares significant sequence similarity with the catalytic domains of the family of serine/threonine protein kinases. Using both genetic and biochemical approaches, the following will be analyzed: (1) the functional role of myristoylation on Vps15p function and localization, (2) the subcellular location and possible site of action of the Vps15 protein, and (3) the presumed kinase activity of the VPS15 gene product. It is anticipated that these studies will result in a detailed view of the organization and control of vesicular traffic between the Golgi and the vacuole, and possibly other inter-organelle transport events in eukaryotic cells. The process of selective recognition, sorting and transport of proteins from one intracellular organelle to another via vesicular carriers represents an essential feature of all eukaryotic cells. The precise mechanisms and machinery that direct these processes are not yet known. This laboratory has isolated a large collection of yeast mutants that exhibit severe defects in protein delivery to the lysosome-like vacuole. The defective genes in these mutants are likely to encode components of the cell's protein sorting apparatus. Toward an understanding of the molecular and biochemical basis of the vacuolar protein sorting ("vps") defects in these mutants, this research will characterize the roles certain of the VPS genes and their products play in the sorting and delivery of vacuolar enzymes from the Golgi complex to the vacuole in yeast. The fundamental similarities between yeast and other eukaryotic cells in their pathways for protein delivery, together with the powerful genetics and molecular approaches available in yeast, make yeast an ideal organism for addressing these problems.

本研究的重点是酵母液泡蛋白分选（VPS）基因之一VPS15。VPS15基因的预测产物与丝氨酸/苏氨酸蛋白激酶家族的催化结构域具有显著的序列相似性。使用遗传学和生化方法，将分析以下内容：(1)肉豆肉酰化对Vps15p功能和定位的功能作用，(2)Vps15蛋白的亚细胞位置和可能的作用位点，以及(3)Vps15基因产物的推定激酶活性。预计这些研究将导致对高尔基体和液泡之间的囊泡运输的组织和控制的详细看法，以及真核细胞中可能的其他细胞器间运输事件。蛋白质通过囊泡载体从一个细胞器到另一个细胞器的选择性识别、分类和运输过程是所有真核细胞的基本特征。指导这些过程的确切机制和机制尚不清楚。本实验室已分离出大量酵母突变体，这些突变体在向溶酶体样液泡输送蛋白质方面表现出严重缺陷。这些突变体中有缺陷的基因很可能编码细胞蛋白质分选装置的组成部分。为了了解这些突变体中液泡蛋白分选（vps）缺陷的分子和生化基础，本研究将描述某些vps基因及其产物在酵母高尔基复合体中液泡酶的分选和传递过程中所起的作用。酵母和其他真核细胞在蛋白质传递途径上的基本相似之处，加上酵母中强大的遗传学和分子方法，使酵母成为解决这些问题的理想生物。