CAA: Cloning and Initial Molecular Characterization of Yeast VPS47 Gene

小行星9628802 这是一个为女性科学家和工程师设立的职业发展奖。 该项目的重点是蛋白质的选择性识别，分选和运输到酵母的溶酶体（液泡），酿酒酵母。 在高等真核生物中，酵母的溶酶体空泡是其降解细胞器，并且几种水解酶通过分泌途径定位在那里。 不同实验室的各种突变体筛选策略已经导致分离出大量的酵母突变体，这些突变体在蛋白质递送到液泡方面表现出严重缺陷，称为液泡蛋白分选突变体（vps突变体）。 这些突变体中的缺陷基因可能编码细胞蛋白质分选装置的成分。 利用改良的筛选方法，本实验室分离出一种新的vps突变体vps47，它代表了一种独特的遗传缺陷：它是迄今为止唯一一种严重的温度敏感vps突变体，在液泡的完整性，结构或功能方面没有缺陷。 这代表了液泡定位机制中一个非常有趣的基因。 该项目将在休假年进行，需要克隆VPS 47基因并确定其特征。 从这些特征中可以深入了解这种和其他vps基因产物的功能。 ***

9628802 Gharakhanian This is a Career Advancement Award for Women Scientists and Engineers. The project focuses on the selective recognition, sorting and transport of proteins to the lysosome (vacuole) of the yeast, Saccharomyces cerevisiae. As in higher eukaryotes, the lysosomal vacuole of the yeast is its degradative organelle, and several hydrolytic enzymes are localized there via the secretory pathway. Various mutant screening strategies in different laboratories have resulted in the isolation of a large collection of yeast mutants that exhibit severe defects in protein delivery to the vacuole, called vacuolar protein sorting mutants (vps mutants). The defective genes in these mutants are likely to encode components of the cell's protein sorting apparatus. Using a modified screening method, this laboratory isolated a new vps mutant, vps47, which represents a unique genetic defect: it is the only severe temperature sensitive vps mutant to date that shows no defects in the integrity, structure, or function of the vacuole. This represents a very interesting gene within the vacuolar localization machinery. This project, which will be carried out during a sabbatical year, entails the cloning and characterization of the VPS 47 gene. Insight into the function of this and other vps gene products will be gained from these characterizations. ***