Role for Protein Phosphorylation in Yeast Vacuolar Protein Sorting

本研究主要针对酵母液泡蛋白中的一种 分选（VPS）基因，VPS 15. VPS 15的预测产品 基因共享显着的序列相似性与催化 丝氨酸/苏氨酸蛋白激酶家族的结构域。 使用 遗传和生物化学方法，以下将是 分析：（1）豆蔻酰化对Vps 15 p的功能作用 功能和定位，（2）亚细胞定位， Vps 15蛋白的可能作用位点，以及（3）推测的 VPS 15基因产物的激酶活性。 预计各国 这些研究将使我们对本组织有一个详细的了解， 以及控制高尔基体和液泡之间的囊泡运输， 以及真核生物中可能的其他细胞器间运输事件 细胞 选择性识别、分类和运输的过程 蛋白质从一个细胞内细胞器到另一个细胞内细胞器， 载体代表了所有真核细胞的基本特征。 指导这些过程的精确机制和机器 目前还不清楚。 这个实验室已经分离出大量的 酵母突变体在蛋白质传递方面表现出严重缺陷 溶酶体样空泡。 这些突变体中的缺陷基因 很可能编码细胞蛋白质分类的成分 设备. 为了理解分子和 液泡蛋白分选（“VPS”）缺陷的生物化学基础 在这些突变体中，这项研究将描述某些 的VPS基因和他们的产品在排序和 将液泡酶从高尔基复合体传递到液泡 在酵母中。 酵母和其他酵母之间的基本相似之处 真核细胞在其蛋白质递送途径中， 利用强大的遗传学和分子方法， 酵母，使酵母成为解决这些问题的理想生物。

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.