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

初级纤毛是细胞的触角，伸出细胞外，介导关键的信号通路。纤毛由一个微管轴组成，它植根于基体，并被一个特殊的膜包围，该膜集中了受体，以正确地解释各种细胞外信号。纤毛缺陷可导致多种疾病。这些疾病包括多囊肾病、视网膜变性和先天性心脏缺陷，而这种细胞器的丧失可能是癌症进展过程中的一个关键事件。******我研究的长期目标是了解纤毛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确定，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发挥其功能。当前提案的目的是了解小说的蛋白质网络- COMMD /洗这种货物从膜在纤毛的形成有关。我们在一项大型蛋白质组学研究中发现了这种复合物，我们有证据表明它与已知与新生纤毛膜相关的基底体成分有关。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来研究：1)该复合体的哪些成分是调解所必需的；(2)哪一个具体的调解步骤受这些因素的影响；3)该络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，在分子水平上理解，在纤毛形成过程中，基底体的蛋白质是如何与膜界面的，从而产生一个功能齐全的纤毛膜。在此过程中，我们将提供基本的见解，不仅有助于理解细胞生长和信号传导，而且有助于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，学习基本的细胞和分子遗传学方法，以及蛋白质组学和功能基因组学的前沿新方法。这种培训将推动下一代加拿大科学家的职业发展。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. *****