MOLECULAR GENETICS OF A LENS INTRINSIC MEMBRANE PROTEIN

晶状体固有膜蛋白的分子遗传学

• 批准号: 2711187
• 负责人: ROBERT L CHURCH
• 金额: $ 21.19万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-08-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2711187
• 关键词: DNA footprinting; cataract; cell differentiation; fiber cell; gel mobility shift assay; gene expression; genetic regulatory element; genetically modified animals; laboratory mouse; lens proteins; membrane proteins; methylation; molecular genetics; transcription factor

DESCRIPTION: (Investigator's Abstract). The ocular lens is an avascular, semisolid organ whose function is to refract incoming light upon the retina. Concomitant with this function is a necessity for the lens to remain transparent. A loss of transparency of the lens is usually permanent and is termed a cataract. Cell and organ functions, such as obtaining nutrients necessary for maintaining the healthy, transparent lens state, for handling of cell waste removal, and for maintenance of overall membrane stability are carried out through select and specific proteins or groupings of proteins within the lens cell membrane. Proper controls on expression of these proteins and their subsequent function is essential for lens survival, and maintenance of transparency. The mechanisms regulating these membrane proteins in the lens are presently unknown. The goal of this project is to understand the regulation of the lens fiber cell membrane protein MP19 gene (LIM2). To achieve this goal, the P.I. will investigate the following topics: a). To characterize the mouse To3 cataract mutant and the human congenital nuclear cataract and their relationship to the lens membrane LIM2 gene. Using transgenic mouse technology to mimic the To3 cataract in order to study the function of MP19. b). To determine the region(s) responsible for transcriptional activity of the human LIM2 gene (cis elements), using CAT reporter gene constructs in transient transfection assays. c). Identify protein:DNA interactions (trans factors) associated with the LIM2 basal promoter region and upstream cis elements using gel mobility shift and DNA foot printing procedures. d). To investigate the methylation patterns of LIM2 upstream regions to determine if methylation of critical cis elements serves to control LIM2 gene function. These studies will give a better understanding of the role of MP19 in the maintenance of lens transparency and its possible role in cataract formation.

描述：(调查员摘要)。人工晶状体是一种 无血管的半固态器官，其功能是折射入射光 在视网膜上。与此函数相伴随的是 镜头保持透明。镜片透明度的损失是 通常是永久性的，被称为白内障。细胞和器官的功能， 比如获取维持健康所需的营养， 透明透镜状态，用于处理电池废物的去除，以及用于 通过选择来维护膜的整体稳定性 和晶状体细胞内的特定蛋白质或蛋白质组 薄膜。适当控制这些蛋白的表达和它们的 后续功能对晶状体的存活和维持至关重要。 透明度。这些膜蛋白在细胞内的调节机制 镜片目前还不得而知。这个项目的目标是了解 晶状体纤维细胞膜蛋白MP19基因(LIM2)的调控。 为了实现这一目标，私人情报局将调查以下主题：a)。 小鼠TO-3突变型白内障与人类先天性白内障的特征 核性白内障及其与晶状体膜LIM2基因的关系 利用转基因小鼠技术模拟TO3白内障 研究了MP19的功能。b)。确定地区(S)责任 对于人类LIM2基因(顺式元件)的转录活性，使用 瞬时转染实验中CAT报告基因的构建。c)。 识别蛋白质：DNA相互作用(反式因子)与 凝胶迁移率测定LIM2基因启动子区和上游顺式元件 移位和DNA脚印程序。d)。为了调查 LIM2上游区域甲基化模式以确定是否 关键顺式元件甲基化调控LIM2基因 功能。 这些研究将使我们更好地理解MP19在体内的作用。 晶状体透明度的维持及其在白内障中的可能作用 队形。