Polycystins and cilia in C. elegans

描述（由申请人提供）：本研究的目的是了解纤毛的形成和功能。秀丽隐杆线虫是研究纤毛生物学的重要模式系统。纤毛的形成、维持和功能需要许多蛋白质。秀丽线虫与人类肾脏疾病有关，包括常染色体显性多囊肾病（ADPKD）。ADPKD影响1/1000个体，通常导致终末期肾病。在人类中，多囊蛋白-1（PC-1）或多囊蛋白-2（PC-2）纤毛机械感觉复合体的突变导致ADPKD。In C.在线虫中，多囊蛋白LOV-1和PKD-2定位于睫状膜上，并且是感觉转导所需的。因此，多囊蛋白和纤毛之间的联系似乎是一个古老的。无论是人肾上皮细胞还是蠕虫的感觉神经元，多囊蛋白最终必须定位于纤毛，以进行细胞的感觉功能。多囊蛋白和其他纤毛蛋白如何定位并进入纤毛（一种空间受限的细胞器）尚不清楚。In C.因此，有可能在活体动物中鉴定控制多囊蛋白纤毛定位和功能的机制。本研究将使用经典和反向遗传学、分子生物学、转基因线虫、显微镜、电生理学、钙成像和生物化学方法来了解多囊蛋白的调节、功能和定位。一个具体的目标是确定和表征的分子机制，调节PKD-2/PC-2纤毛蛋白的定位和功能。第二个目的是研究驱动蛋白KLP-6在纤毛蛋白运输和感觉中的作用，以及鉴定KLP-6货物。第三个目标是鉴定纤毛发生和多囊蛋白纤毛定位所需的新基因。这些研究在一个易处理的模式生物将提供基本的见解纤毛的形成和功能的机制。

DESCRIPTION (provided by applicant): The goal of this research is to understand how cilia form and function. The nematode Caenorhabditis elegans is a powerful model system to study cilia biology. Many proteins that are required for formation, maintenance, and function of cilia in C. elegans are linked to human renal diseases, including autosomal dominant polycystic kidney disease (ADPKD). ADPKD affects 1 in 1000 individuals, often resulting in end-stage renal disease. In humans, mutations in the polycystin-1 (PC-1) or polycystin-2 (PC-2) ciliary mechanosensory complex cause ADPKD. In C. elegans, the polycystins LOV-1 and PKD-2 localize on the ciliary membrane and are required for sensory transduction. Hence, the connection between the polycystins and cilia seems to be an ancient one. The polycystins must ultimately be localized to cilia in order to conduct the sensory function of the cell, whether it is a human renal epithelial cell or a worm sensory neuron. How the polycystins and other ciliary proteins localize and gain access to the cilium, a spatially restricted organelle, is not known. In C. elegans it is possible to identify the mechanisms controlling polycystin ciliary localization and function in living animals. This study will use classical and reverse genetics, molecular biology, transgenic nematodes, microscopy, electrophysiology, calcium imaging, and biochemical methods to understand the regulation, function, and localization of the polycystins. One specific aim is to identify and characterize the molecular mechanisms regulating PKD-2/PC-2 ciliary protein localization and function. A second aim is to examine the role of the kinesin KLP-6 in ciliary protein transport and sensation, as well as identify KLP-6 cargoes. A third aim is to identify new genes required for ciliogenesis and polycystin ciliary localization. These studies in a tractable model organism will provide basic insights into the mechanisms governing cilia formation and function.