PRESENILIN GENE STUDY IN DROSOPHILA MELANOGASTER AS MODEL

以果蝇为模型的早老素基因研究

• 批准号: 6485268
• 负责人: ROBERT F CLARK
• 金额: $ 17.91万
• 依托单位: MEHARRY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2002-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6485268
• 关键词: Alzheimer's disease; Drosophilidae; developmental genetics; disease /disorder onset; gene interaction; gene mutation; genetic models; genetic transcription; nucleic acid sequence; presenilin; Alzheimer's disease; Drosophilidae; developmental genetics; disease /disorder onset; gene interaction; gene mutation; genetic models; genetic transcription; nucleic acid sequence; presenilin

Genetic linkage studies in early onset-familial Alzheimer's disease (AD) families have determined that mutations in at least three loci will cause the disease: the amyloid beta-protein precursor gene, the presenilin-1 gene (PS-1), and the presenilin-2 gene (PS-2). While it has been reported that beta-amyloid production is abnormal in fibroblasts from families carrying PS-1 and PS-2 mutations, and a homologues protein in C. elegans affects Notch signaling, the function of the presenilin gene is presently unknown. Therefore, the long-term objective of this application is to determine the function of the presenilin protein and other presenilin- interacting proteins. Drosophila melanogaster is an ideal model organism for determining the function of uncharacterized genes, which in this case is made even more ideal as most studies of Notch signaling have been carried out in Drosophila. Using the high conservation among conservation among presenilin genes, our lab has successfully coned the gene in Drosophila. The first specific aim of this project will be to characterize this gene in Drosophila, studying its gene structure, its developmental transcription pattern, and its localization in embryos, larvae, pupae, and adult flies, DNA sequencing. Southern and Northern blotting techniques, and light microscopy will be used to carry out this part of the project. The second specific aim will be to produce null mutations of this gene and study their effects in Drosophila. P-element insertion mutagenesis will be used to produce the mutant flies, and the flies will be characterized genetically. The third specific aim will be to determine which genes interact with the presenilin gene. Fly genetics and yeast two-hybrid approaches will be used to find these presinilin- interacting genes. This project will lead to a greater understanding about the biology and function of the presenilin genes, the major cause of early- onset familial Alzheimer's disease. By determining the functions of the presenilins, and their interactions with other genes, a greater understanding of the pathogenesis of AD will be gained, which may lead eventually to improved treatment of the disease. Additionally, a long- term study of these presenilin-interacting proteins will help us to identify other human loci which may cause or influence AD in both early-onset and late-onset cases.

早期发作 - 家庭阿尔茨海默氏病（AD）家族中的遗传连锁研究已经确定至少三个基因座的突变会引起该疾病：淀粉样蛋白β-蛋白蛋白前体基因基因，Presenilin-1基因（PS-1）和Presenilin-2基因（PS-2）。据报道，β-淀粉样蛋白的产生在携带PS-1和PS-2突变的家族的成纤维细胞中异常，而秀丽隐杆线虫中的同源物蛋白会影响Notch信号传导，但Presenilin Gene的功能目前尚不清楚。因此，该应用的长期目标是确定presenilin蛋白和其他presenilin-相互作用蛋白的功能。果蝇Melanogaster是确定未表征基因功能的理想模型生物，在这种情况下，由于大多数Notch信号的研究已在果蝇中进行。我们的实验室在Presenilin基因中使用了高保守性，在果蝇中成功地将该基因固定在果蝇中。该项目的第一个具体目的是在果蝇中表征该基因，研究其基因结构，其发育转录模式以及其在胚胎，幼虫，pupe虫和成年果蝇中的定位，DNA测序。南部和北印迹技术以及光学显微镜将用于执行该项目的这一部分。第二个特定目的是产生该基因的无效突变，并研究其在果蝇中的作用。 P元素插入诱变将用于产生突变蝇，并将果蝇从遗传上进行表征。第三个特定目的是确定哪些基因与presenilin基因相互作用。 Fly遗传学和酵母双杂交方法将用于找到这些相互作用的基因。该项目将使人们对Presenilin基因的生物学和功能有更深入的了解，这是早期发作家族性阿尔茨海默氏病的主要原因。通过确定寄生虫的功能及其与其他基因的相互作用，将获得对AD发病机理的更多了解，这可能最终导致对疾病的治疗。此外，对这些老年蛋白相互作用蛋白的长期研究将有助于我们确定可能在早发和晚发病例中引起或影响AD的其他人类基因座。