Dissecting the molecular function of basigin a protein required for normal struct

描述(由申请人提供)：眼睛的退行性疾病，特别是视网膜外营养不良，是导致无法治愈的失明的主要原因。一个可能在视网膜病变中起作用的基因是basigin基因(在人类中也称为EMMPRIN)。Basigin是编码细胞表面免疫球蛋白的基因家族的成员，该蛋白与细胞间的相互作用有关。我们以果蝇为模型系统，研究basigin在基本视觉系统、细胞生物学和视觉系统功能中的作用。我们已经用苍蝇确定了视觉系统中的几种basigin功能，其中许多功能与脊椎动物中描述的功能一致。例如，basigin突变的小鼠表现出异常的视网膜电信号(ERG)，就像眼睛中basigin功能突变的马赛克苍蝇一样(Clip等人，2003；Curtin等人，2007)。Basigin突变小鼠表现出光感受器(PC)细胞结构的改变(Ochreitor等人，2001年)。果蝇basigin是正常光感受器细胞结构所必需的(Curtin等人，2005年)。Basigin参与了鸟类视觉系统分离细胞中神经元-神经胶质细胞的相互作用(Fadool和Linser，1993)。果蝇basigin是视板神经元-神经胶质细胞相互作用所必需的(Curtin等人，2007年)。所有这些巧合有力地证明，我们所了解的Basigin在果蝇视网膜中的分子功能将有助于我们了解其在哺乳动物视网膜中的功能。最后，苍蝇和老鼠basigin都可以修复马赛克basigin苍蝇视觉系统中的缺陷，这表明哺乳动物basigin可以与果蝇basigin进行类似的分子接触(Curtin等人，2005,2007)。我们的主要目标是了解basigin促进正常视觉系统结构和功能的分子机制。我们的初步数据显示，basigin形成了蛋白质复合体。我们正在使用一种酵母双杂交试验，旨在检测细胞膜上的蛋白质相互作用，以确定可能与basigin相互作用的蛋白质和编码它们的基因。我们将利用遗传/功能研究和生物化学来确定这些蛋白质相互作用的功能意义，以及探索特定的basigin蛋白修饰对basigin细胞功能的意义。能够独立于动物其他部位专门操纵眼睛的遗传构成，为我们提供了一个特殊的系统来研究basigin的分子功能和basigin相互作用蛋白的基因。 公共卫生相关性：拟议的研究通过探索basigin/EMMPRIN在视觉系统的维持和功能中的分子作用，与人类健康相关。该项目的主要目的是识别眼睛中与basigin相互作用的蛋白质，并分析这些蛋白质的编码基因，以便我们更好地了解basigin如何影响视觉系统的发育和功能。

DESCRIPTION (provided by applicant): Degenerative diseases of the eye, specifically outer retinal dystrophies, are a major cause of incurable blindness. One gene that may play a role in retinopathy is the basigin gene (aka EMMPRIN in humans). Basigin is a member of a gene family that encodes cell surface IgG proteins that are implicated in cellular interactions. We are using Drosophila as a model system to study basigin's role in basic visual system cell biology and visual system function. We have identified several basigin functions in the visual system using flies, many of which are co-incident with functions described in vertebrates. For example, basigin mutant mice show abnormal electroretinograms (ERGs) as do mosaic flies in which basigin function is mutant in the eye (Clamp et al., 2003; Curtin et al., 2007). Basigin mutant mice show altered photoreceptor (PC) cell structure (Ochreitor et al., 2001). Drosophila basigin is required for normal photoreceptor cell structure (Curtin et al., 2005). Basigin has been implicated in neuron-glia interaction in dissociated cells from the avian visual system (Fadool and Linser, 1993). Drosophila basigin is necessary for neuron-glia interactions in the optic lamina (Curtin et al., 2007). All these co-incidences argue strongly that what we learn about basigin molecular function in the Drosophila retina will help us understand its function in the mammalian retina. Lastly, both fly and mouse basigin can rescue the defects seen in the visual system of mosaic basigin flies showing that mammalian basigin can make similar molecular contacts as Drosophila basigin (Curtin et al., 2005, 2007). Our main goal is to understand the molecular mechanisms by which basigin acts to promote normal visual system structure and function. Our preliminary data show that basigin forms protein complexes. We are using a yeast two-hybrid assay designed to detect protein interactions at the membrane to identify putative basigin-interacting proteins and the genes that encode them. We will use genetic/ functional studies and biochemistry to identify the functional significance of these protein interactions as well as exploring the significance of specific basigin protein modifications for basigin cellular functions. The ability to specifically manipulate the genetic make-up of the eye independent of the rest of the animal provides us with an exceptional system to study the molecular function of basigin and the genes for basigin interacting proteins. PUBLIC HEALTH RELEVANCE: The proposed studies have relevance to human health by exploring the molecular role of basigin/EMMPRIN in the maintenance and function of the visual system. The project's main aim is to identify basigin-interacting proteins in the eye and to analyze the genes that code for these proteins so that we can better understand how basigin affects visual system development and function.