MOLECULAR GENETICS OF A LENS INTRINSIC MEMBRANE PROTEIN

描述：（研究者摘要）。 目镜透镜是一种 一种无血管的半固体器官，其功能是吸收入射光 在视网膜上。 与此功能相伴随的是， 透镜保持透明。 透镜的透明度损失是 通常是永久性的，称为白内障。 细胞和器官功能， 例如获得维持健康所必需的营养素， 透明透镜状态，用于处理细胞废物去除，以及用于 整体膜稳定性的维持通过选择 以及透镜细胞内的特定蛋白质或蛋白质群 膜的 适当控制这些蛋白质的表达及其 随后的功能对于透镜的存活和维持是必不可少的， 透明度 这些膜蛋白的调节机制在细胞膜上的表达与细胞膜的形成有关。 透镜目前是未知。 这个项目的目标是了解 透镜纤维细胞膜蛋白MP19基因（LIM2）的调控。 为了实现这一目标，P.I.将研究以下主题：a）。 为了表征小鼠To3白内障突变体和人类先天性白内障突变体， 核性白内障及其与透镜膜LIM2基因的关系。 利用转基因小鼠技术模拟To3白内障， 研究MP19的功能。 B）。确定负责区域 对于人LIM2基因（顺式元件）的转录活性，使用 瞬时转染测定中的CAT报告基因构建体。 （c）。 确定蛋白质：DNA相互作用（反式因子）与 使用凝胶迁移率的LIM2基础启动子区和上游顺式元件 转移和DNA足迹程序。 d）。探讨 LIM2上游区域的甲基化模式，以确定 关键顺式元件的甲基化控制LIM2基因 功能 这些研究将使我们更好地了解MP19的作用， 透镜透明度的维持及其在白内障中的可能作用 阵

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.