Understanding the molecular mechanisms of cilia formation

了解纤毛形成的分子机制

• 批准号: 10713810
• 负责人: Tomoharu Kanie
• 金额: $ 36.25万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2028-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10713810
• 关键词: Cell Cycle; Cell membrane; Cells; Cellular Membrane; Cellular Structures; Centrioles; Centrosome; Cilia; Disease; Distal; Embryonic Development; Environment; FREQ gene; Functional disorder; G1 Phase; Health Sciences; Homeostasis; Human; Lipids; Mitosis; Molecular; Monitor; Monomeric GTP-Binding Proteins; Mothers; Movement; Obesity; Oklahoma; Organelles; Pathogenesis; Play; Proteins; Research Project Grants; Retinal Degeneration; Role; Signal Transduction; Structure; Syndrome; Universities; Vesicle; base; biochemical tools; ciliopathy; embryo tissue; extracellular; myristoylation; receptor; recruit

Project Summary: The Primary cilium is an organelle that majority of human cells possess with only one per a cell. This immotile cellular structure senses extracellular signaling via receptors that specifically accumulate on the ciliary portion of the cellular membrane. Dysfunction in the structure or the function of the organelle results in pleiotropic disorders called ciliopathies, which include obesity and retinal degeneration. The cilium needs to be disassembled prior to mitosis, as cells use centrosome, from which the cilium extends, as spindle poles. Cells reassemble the cilium in G0/G1 phase of the cell cycle. My lab at University of Oklahoma Health Science Center interrogates the molecular mechanisms of the formation of the cilium. The initial step of the cilium formation is recruitment of a ciliary vesicle, to the distal end of the mother centriole. The small vesicles fuse to form a larger vesicle, which finally merges with plasma membrane to form the primary cilium. While the attachment of the ciliary vesicle to the centriole is the first step, many important questions remain to be elucidated. One potential problem that has hampered our understanding of the ciliary vesicle is the lack of markers of the ciliary vesicle. To overcome this challenge, I previously characterized a small GTPase, RAB34, a more specific marker for the ciliary vesicle, and determined that Neuronal Calcium Sensor-1 (NCS1) captures the ciliary vesicle using its myristoylation motif at the ciliary base. Using these proteins as powerful biochemical tools, we will focus on answering key unsolved questions including the composition and origin of the ciliary vesicle. We will isolate the ciliary vesicle to uncover the lipid and protein composition of the vesicle, monitor the movement of the ciliary vesicle from its emergence to reveal the origin of the vesicle, and screen for the regulators that determine how the vesicle is generated and transported to the centriole. If successful, our research projects will uncover the largely unexplored molecular mechanisms of the early step of the cilium formation, which will pave the way to get to the root of the pathogenesis of the syndrome caused by ciliary dysfunction.

项目概要： 初级纤毛是一种细胞器，大多数人体细胞都具有，每个细胞只有一个。 这种不动的细胞结构通过受体感知细胞外信号， 聚集在细胞膜的纤毛部分。结构功能障碍或 细胞器的功能导致称为纤毛病的多效性疾病，包括 肥胖和视网膜变性。纤毛在有丝分裂前需要被分解， 以纤毛延伸的中心体作为纺锤体的极。细胞重新组装纤毛， 细胞周期的G 0/G1期。我在俄克拉荷马州大学健康科学中心的实验室 探讨纤毛形成的分子机制。的初始步骤 纤毛的形成是纤毛囊泡向中心粒末端的募集。的 小的囊泡融合形成更大的囊泡，最后与质膜融合形成 初级纤毛纤毛囊附着在中心粒上是第一步， 许多重要问题仍有待阐明。一个潜在的问题阻碍了 我们对睫状泡的理解是缺乏睫状泡的标记物。克服 在这一挑战中，我以前描述了一个小的GTdR，RAB 34，一个更特异的标记物， 纤毛囊泡，并确定神经元钙传感器-1（NCS 1）捕获纤毛囊泡， 囊泡利用其在纤毛基部的肉豆蔻酰化基序。利用这些蛋白质作为强大的 生物化学工具，我们将专注于回答关键未解决的问题，包括组成 和睫状囊的起源。我们将分离出睫状囊泡以揭示脂质， 蛋白质组成的囊泡，监测纤毛囊泡的运动，从其 出现，以揭示囊泡的起源，并筛选决定如何 产生囊泡并运输到中心粒。如果成功，我们的研究项目 将揭示纤毛早期阶段的分子机制， 从而为从根本上探讨本病的发病机理奠定基础 是由纤毛功能障碍引起的