Identifying Gene Functions at Late Endosome and Lysosome in Yeast

描述（由申请人提供）：分子的正确分选和囊泡运输至降解溶酶体是所有真核细胞的基本特征。酵母酿酒酵母的晚期内体和液泡在功能上等同于哺乳动物晚期内体和溶酶体;其液泡递送的羧肽酶Y（CPY）途径与哺乳动物细胞中的甘露糖-6-磷酸途径平行。酵母提供了传统和分子遗传学工具的优势;在溶酶体运输中鉴定的绝大多数酵母基因在人类中具有直系同源物。对液泡运输和功能相关基因的两个单独的全基因组筛选的结果使我们发现：1）几个以前未表征的在内体和液泡界面起作用的基因（ENV基因），2）一小部分参与液泡运输和功能的基因（HHY基因），当缺失时，这些基因赋予潮霉素B严重超敏性。此外，所有hhy突变体还显示出指示缺陷性TOR激酶信号传导的药物超敏性，TOR激酶信号传导是通过营养物和生长因子传感的细胞生长和增殖的主要调节剂。在这种竞争性的更新，我们提出了多方面的表征新基因ENV 9具有显着的身份，人类视黄醇脱氢酶RDH 12-一个基因牵连在早发性视网膜变性。我们还建议通过酵母遗传学、分子和显微镜方法探索空泡运输、TOR激酶信号传导和潮霉素B超敏反应的界面。在这个区域的建议固有的，是一个少数民族博士后研究人员的目的继续发展，并继续在一个全面的西班牙裔服务研究所本科生和硕士生的生产性研究培训。溶酶体蛋白酶和货物的错误定位与溶酶体贮积病和阿尔茨海默病（AD）相关。特别是在运输的晚期内体到溶酶体阶段的缺陷已经成为青少年和老年神经变性疾病中可能的潜在机制。TOR激酶信号传导的过度活化与良性和恶性肿瘤发生以及包括糖尿病和肥胖症在内的代谢疾病相关。 公共卫生相关性：拟议的研究与青少年和迟发性神经变性相关的公共卫生问题有关，分别表现在溶酶体贮积病和阿尔茨海默病中。它还与与肿瘤发生和代谢疾病（包括糖尿病和肥胖）相关的公共卫生问题相关。

DESCRIPTION (provided by applicant): Correct sorting and vesicular trafficking of molecules to the degradative lysosome is an essential feature of all eukaryotic cells. The late endosome and vacuole of the yeast Saccharomyces cerevisiae are functionally equivalent to the mammalian late endosome and lysosome; its carboxy peptidase Y (CPY)-pathway of vacuolar delivery parallels the mannose-6-phosphate pathway in mammalian cells. Yeast offers the advantage of both conventional and molecular genetics tools; large majority of yeast genes identified in lysosomal trafficking have orthologues in humans. Results of two separate genome wide screens for vacuolar trafficking and function related genes have led us to 1)several previously uncharacterized genes that have a role at endosome and vacuole interface (ENV genes), 2) a small set of genes involved in vacuolar trafficking and function that confer severe hypersensitivity to hygromycin B when deleted (HHY genes). Furthermore, all hhy mutants also showed drug hypersensitivities indicative of defective TOR kinase signaling, a master regulator of cell growth and proliferation through nutrient and growth factor sensing. In this competitive renewal, we propose multifaceted characterization of the novel gene ENV9 which has significant identity to human retinol dehydrogenase RDH12 - a gene implicated in early onset retinal degeneration. We also propose to explore the interface of vacuolar trafficking, TOR kinase signaling, and hygromycin B hypersensitivity by yeast genetics, molecular, and microscopic approaches. Inherent in this AREA proposal, is the aim to continue development of a minority post-doctoral researcher and continue productive research training of undergraduate and Master's students within a comprehensive Hispanic Serving Institute. Mislocalization of lysosomal proteases and cargo is associated with Lysosomal Storage Diseases and Alzheimer's disease (AD). Defects specifically at the late endosome to lysosome stage of trafficking have emerged as the possible underlying mechanism in both juvenile and aging neurodegeneration diseases. Hyper activation of TOR kinase signaling is associated with benign and malignant tumorigenesis, as well as with metabolic diseases including diabetes and obesity. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health issues associated with both juvenile and late onset neurodegeneration as manifested in Lysosomal Storage Diseases and Alzheimer's disease, respectively. It is also relevant to public health issues associated with tumorigenesis and metabolic diseases including diabetes and obesity.