Regulation of gap junctions by stomatin-like proteins

描述（由申请人提供）：缝隙连接是普遍存在的细胞间通道，在发育和生理学中发挥非常重要的作用。然而，很少有人知道的蛋白质，可能相互作用和调节间隙连接。无脊椎动物系统可用于鉴定进化上保守的间隙连接调节剂，因为主要结构特征和功能性质在无脊椎动物和脊椎动物间隙连接之间是保守的。In C.在线虫中，基因UNC-7、UNC-9、UNC-1和UNC-24的独立功能丧失突变引起相似的表型，包括运动缺陷和对挥发性麻醉剂的敏感性改变。UNC-7和UNC-9编码作为间隙连接蛋白的连接蛋白，而UNC-1和UNC-24编码气孔蛋白样蛋白。尚不清楚为什么连接蛋白和气孔蛋白样蛋白的功能障碍导致相似的表型。该提议的目的是检验以下假设，即α-1和α-24是间隙连接的调节剂。具体目的是（1）探索口蛋白-9和口蛋白样蛋白之间相互作用的分子基础，（2）确定口蛋白-1和口蛋白-24是否与口蛋白-9的羧基末端相互作用以调节间隙连接门控，以及（3）确定口蛋白-1、口蛋白-24、口蛋白-7和口蛋白-9是否在神经元中相互作用以调节运动。各种电生理学、遗传学和细胞生物学方法将被用来分析这些蛋白质之间的相互作用。elegans以及哺乳动物异源表达系统。该项目的长期目标是了解间隙连接调节的进化保守机制，并确定用于治疗和管理间隙连接相关疾病的候选药物靶点。

DESCRIPTION (provided by applicant): Gap junctions are ubiquitous intercellular channels that play very important roles in development and physiology. However, little is known about proteins that may interact with and modulate gap junctions. Invertebrate systems could be used to identify evolutionarily conserved gap junction modulators because major structural features and functional properties are conserved between invertebrate and vertebrate gap junctions. In C. elegans, independent loss-of-function mutations of the genes unc-7, unc-9, unc-1, and unc-24 cause similar phenotypes, including locomotion defects and altered sensitivity to volatile anesthetics. unc-7 and unc-9 encode innexins, which are gap junction proteins, whereas unc-1 and unc-24 encode stomatin-like proteins. It is unknown why dysfunctions of the innexins and stomatin-like proteins cause similar phenotypes. The purpose of this proposal is to test the hypothesis that UNC-1 and UNC-24 are modulators of gap junctions. The specific aims are (1) to explore molecular basis for interactions between UNC-9 and stomatin-like proteins, (2) to determine whether UNC-1 and UNC-24 interact with the carboxyl terminus of UNC-9 to modulate gap junction gating, and (3) to determine whether UNC-1, UNC-24, UNC-7, and UNC-9 interact in neurons to modulate locomotion. A variety of electrophysiological, genetic, and cell biological approaches will be used to analyze interactions among these proteins in C. elegans as well as in a mammalian heterologous expression system. The long term goals of this project are to understand evolutionarily conserved mechanisms of gap junction regulation and to identify candidate drug targets for treatment and management of gap junction-related diseases.