Understanding membrane shape remodelling in cilia formation and function

了解纤毛形成和功能中的膜形状重塑

• 批准号: BB/X013030/1
• 负责人: Mark Van Breugel
• 金额: $ 60.58万
• 依托单位: Queen Mary University of London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FX013030%2F1
• 关键词: Understanding; membrane; shape; remodelling; cilia

Lipid membranes provide boundaries in biological systems that enable cells to maintain specialised internal compartments ("cell organelles") with separate functions. However, lipid membranes are not rigid, stable structures, but are dynamic and continuously exchange material. To maintain the shapes of their organelles and allow their function, cells use a whole battery of proteins that remodel lipid membranes. The mechanisms of how this happens are incompletely understood and it is often unclear how important the exact regulation of membrane curvature in these events is.A case in point are cilia, hair-like cell projections that are crucial for animals. Cilia serve as motility devices and antennas through which cells sense their environment. They are formed by centrioles, large barrel-shaped, protein structures with distal appendages (DAs) that radiate outwards from one of their ends. Membrane remodelling is critical for cilia formation. This includes the docking of small membrane spheres (vesicles) against the DAs, their fusion and deformation by the extension of the centriole barrel as well as the maintenance of a highly curved membrane region at the base of the cilium where the DAs (then called "transition fibres") dock the centriole barrel ("basal body"). Dysfunction of many of the involved players results in ciliopathies, chronic diseases that affect an estimated 1:1000 people and have no curative treatment. Additionally, cilia are also implicated in other important human disorders such as cancer and heart diseases. Thus, a better understanding of the link between membrane remodelling and cilia will help the development of approaches to rectify malfunctions of the underlying processes in disease.Crucial questions concerning membrane remodelling in cilia formation and function remain unanswered. While DAs associate with known membrane shaping molecules, it is unclear how they organise and coordinate them in space to bring about the complex cilia formation pathway. Furthermore, DAs make intimate contact to the remodelled membranes in this process, which argues that DAs are more than just temporary recruitment platforms for other proteins that then do the job. However, it is not known whether (and how) the DA components on their own contribute to membrane binding and shaping. There are no purification methods available for DAs that would allow to address these questions. We will develop such a method based on centriole fragmentation and use the purified DAs to visualise their high-resolution architecture and test their activity towards membranes in biochemical assays. This will enable us to understand how DAs work, to develop tools to further dissect their function in cells and to gain insights into how their dysfunction in human disease might be rectified. Furthermore, we will address how important the precise shape of the highly curved membrane region contacted by the DAs/transition fibres is for cilia. The membrane shaping DZIP1-Chibby1-FAM92 complex associates with DAs and localises to this membrane region. Importantly, it functions in both cilia formation and basal body docking strongly arguing that it is an active player in maintaining or sensing the local membrane shape there. Our recent work on this complex shows how we can change its membrane shaping properties through protein engineering. We will use the engineered complex in cells to study how altering its curvature induction affects the local membrane shape as well as the generation of cilia and their function. Together, the proposed work will shed light on an ill-understood but fundamental process in animals and reveal how protein complexes mutated in human ciliopathies function in it. The gained insights, as well as our tools and approaches will open new research avenues in important animal models and facilitate the development of strategies for alleviating dysfunctions of this process in disease.

Lipid membranes provide boundaries in biological systems that enable cells to maintain specialised internal compartments ("cell organelles") with separate functions. However, lipid membranes are not rigid, stable structures, but are dynamic and continuously exchange material. To maintain the shapes of their organelles and allow their function, cells use a whole battery of proteins that remodel lipid membranes. The mechanisms of how this happens are incompletely understood and it is often unclear how important the exact regulation of membrane curvature in these events is.A case in point are cilia, hair-like cell projections that are crucial for animals. Cilia serve as motility devices and antennas through which cells sense their environment. They are formed by centrioles, large barrel-shaped, protein structures with distal appendages (DAs) that radiate outwards from one of their ends. Membrane remodelling is critical for cilia formation. This includes the docking of small membrane spheres (vesicles) against the DAs, their fusion and deformation by the extension of the centriole barrel as well as the maintenance of a highly curved membrane region at the base of the cilium where the DAs (then called "transition fibres") dock the centriole barrel ("basal body"). Dysfunction of many of the involved players results in ciliopathies, chronic diseases that affect an estimated 1:1000 people and have no curative treatment. Additionally, cilia are also implicated in other important human disorders such as cancer and heart diseases. Thus, a better understanding of the link between membrane remodelling and cilia will help the development of approaches to rectify malfunctions of the underlying processes in disease.Crucial questions concerning membrane remodelling in cilia formation and function remain unanswered. While DAs associate with known membrane shaping molecules, it is unclear how they organise and coordinate them in space to bring about the complex cilia formation pathway. Furthermore, DAs make intimate contact to the remodelled membranes in this process, which argues that DAs are more than just temporary recruitment platforms for other proteins that then do the job. However, it is not known whether (and how) the DA components on their own contribute to membrane binding and shaping. There are no purification methods available for DAs that would allow to address these questions. We will develop such a method based on centriole fragmentation and use the purified DAs to visualise their high-resolution architecture and test their activity towards membranes in biochemical assays. This will enable us to understand how DAs work, to develop tools to further dissect their function in cells and to gain insights into how their dysfunction in human disease might be rectified. Furthermore, we will address how important the precise shape of the highly curved membrane region contacted by the DAs/transition fibres is for cilia. The membrane shaping DZIP1-Chibby1-FAM92 complex associates with DAs and localises to this membrane region. Importantly, it functions in both cilia formation and basal body docking strongly arguing that it is an active player in maintaining or sensing the local membrane shape there. Our recent work on this complex shows how we can change its membrane shaping properties through protein engineering. We will use the engineered complex in cells to study how altering its curvature induction affects the local membrane shape as well as the generation of cilia and their function. Together, the proposed work will shed light on an ill-understood but fundamental process in animals and reveal how protein complexes mutated in human ciliopathies function in it. The gained insights, as well as our tools and approaches will open new research avenues in important animal models and facilitate the development of strategies for alleviating dysfunctions of this process in disease.