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.

项目摘要： 主要的纤毛是一个细胞器，大多数人细胞只有一个细胞。 这种immotile的细胞结构通过受体感受细胞外信号传导，特别是 堆积在细胞膜的睫状部分。结构或 细胞器的功能导致多效性疾病称为纤毛病，其中包括 肥胖和视网膜变性。纤毛在有丝分裂之前需要拆卸，因为细胞 使用中心体，将纤毛延伸为主轴杆。细胞重新组装纤毛 细胞周期的G0/G1相。我在俄克拉荷马大学健康科学中心的实验室 询问纤毛形成的分子机制。的第一步 纤毛形成是募集睫状囊泡，直到母中心的远端。这 小囊泡融合形成较大的囊泡，最终与质膜合并形成 原发性纤毛。虽然睫状囊泡在中心位置是第一步，但 许多重要的问题仍有待阐明。一个受阻的潜在问题 我们对睫状囊泡的理解是缺乏睫状囊泡的标记。克服 这个挑战，我以前表征了一个小的GTPase Rab34，这是一个更具体的标记 睫状囊泡，并确定神经元钙传感器1（NCS1）捕获纤毛 使用其肉体底部的肉鸡囊泡在纤毛基部。使用这些蛋白质作为功能 生化工具，我们将专注于回答关键的未解决问题，包括构图 和睫状囊泡的起源。我们将隔离睫状囊泡以发现脂质，并 囊泡的蛋白质组成，监测睫状囊泡的运动 出现以揭示囊泡的起源，以及确定如何确定调节器的屏幕 囊泡是生成并运输到中心的。如果成功，我们的研究项目 将发现纤毛早期步骤的很大程度上未开发的分子机制 形成，这将铺平道路，以达到综合征发病机理的根源 由睫状功能障碍引起。