CAA: Cloning and Initial Molecular Characterization of Yeast VPS47 Gene

9628802 Gharakhanian这是一个女性科学家和工程师的职业进步奖。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)。与高等真核生物一样，酵母菌的溶酶体液泡是其降解细胞器，几种水解酶通过分泌途径定位在那里。不同实验室的不同突变体筛选策略导致了大量酵母突变体的分离，这些突变体在向液泡运送蛋白质的过程中表现出严重的缺陷，称为空泡蛋白分选突变体(VPS突变体)。这些突变体中的缺陷基因很可能编码细胞蛋白质分选装置的组件。通过改进的筛选方法，本实验室分离到一个新的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个显示液泡的完整性、结构或功能没有缺陷的严重温度敏感型VPs突变体。这代表了液泡定位机制中一个非常有趣的基因。该项目将在一年的休假期间进行，需要克隆和鉴定VPS 47基因。我们将从这些特征中深入了解该VPS基因产物和其他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. ***