The role of a cilium assembly subnetwork at the basal body-vesicle membrane interface

初级纤毛是细胞的触角，延伸出细胞并介导关键的信号通路。 纤毛由微管轴组成，其扎根于基体，并被专门的膜包围，该膜集中有受体以正确地解释不同的细胞外信号。睫状体缺损导致多种疾病。 这些包括多囊肾病，视网膜变性和先天性心脏病，这种细胞器的丢失可能是癌症进展过程中的关键事件。我研究的长期目标是了解睫状膜和底层结构是如何组装和维持的。虽然这种细胞器的许多成分已被鉴定，但尚不清楚这些蛋白质如何相互作用或与细胞环境相互作用以实现其功能。 目前的提议旨在了解一种新的蛋白质网络- COMMD/WASH -如何在纤毛形成过程中参与细胞膜的货物分类。 我们在一项大型蛋白质组学研究中发现了这种复合物，并且我们有证据表明它与已知与新生睫状体膜相关的基底体成分相关。 我们也有数据表明它是细胞产生正常纤毛所必需的。 我们将使用基因破坏，超分辨率显微镜和蛋白质组学研究：1）该复合物的哪些组分是纤毛所需的; 2）纤毛的确切步骤受这些组分的影响; 3）该复合物的货物分选功能如何影响纤毛膜。我们将利用这些知识来了解，在分子水平上，在纤毛形成过程中，基体的蛋白质如何与膜接触，从而产生一个功能性的睫状膜。 在这样做的过程中，我们将提供基本的见解，以帮助理解，不仅细胞生长和信号，而且如何更好地治疗人类疾病。 拟议的研究计划将培养三名研究生和多名本科生在基本的细胞和分子遗传学方法，以及在蛋白质组学和功能基因组学的尖端新方法。 这项培训将推动下一代加拿大科学家的职业生涯。 由于这些原因，这项研究对加拿大的健康和知识经济以及整个加拿大人都有长期的好处。*****

Primary cilia are cellular antennae' that extend out of the cell and mediate critical signaling pathways. A cilium consists of a microtubular shaft which is rooted at the basal body, and surrounded by a specialized membrane that is concentrated with receptors to correctly interpret diverse extracellular cues. Ciliary defects lead to large spectrum of diseases. These include polycystic kidney disease, retinal degeneration and congenital heart defects, and the loss of this organelle may be a key event during cancer progression.******The long term objective of my research is to understand how the ciliary membrane and underlying structures are assembled and maintained. While many of the constituents of this organelle have been identified, it is unclear how these proteins interact with each other or with the cellular milieu for their function. The current proposal is designed to understand how a novel network of proteins - COMMD/WASH - that sort cargo from membranes is involved during cilium formation. We found this complex in a large proteomic study, and we have evidence that it associates with components of the basal body known to associate with the nascent ciliary membrane. We also have data indicating that it is required for cells to produce normal cilia. We will use gene disruption, super-resolution microscopy and proteomics to study : 1) which components of this complex are required for ciliation; 2) which exact step of ciliation is affected by these components; 3) how the cargo sorting function of this complex affects the ciliary membrane.******We will leverage this knowledge to understand, at a molecular level, how the proteins at the basal body interface with membranes during cilium formation, so as to produce a functioning ciliary membrane. In doing so, we will provide fundamental insights to help understand, not only cellular growth and signalling, but also how to better treat human disease. The proposed research program will train three graduate students and multiple undergraduate associates in fundamental cell and molecular genetic methods, as well as in cutting-edge new approaches in proteomics and functional genomics. This training will propel the next generation of Canadian scientists in their careers. For these reasons, this research has long-term benefits to Canada's health and knowledge economies, as well as to Canadians at large. *****