Functional interplay of ciliary trafficking complexes and motor proteins.

纤毛运输复合物和运动蛋白的功能相互作用。

• 批准号: BB/S013024/1
• 负责人: David Stephens
• 金额: $ 59.61万
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS013024%2F1
• 关键词: Functional; interplay; ciliary; trafficking; complexes

Primary cilia project from the surface of nearly all human cells to serve as signalling platforms. They are essential for human and animal development and are also required throughout life to control pathways that relate to the formation and maintenance of bone, kidney function, signalling in the brain and many more body functions. These cilia are built and maintained by a series of large multi-protein complexes. Our two labs in the UK and Japan have made significant contributions to our understanding of the key machines involved. Most recently we have each defined the function of two microtubule motor protein complexes, dynein-2 and kinesin-2 in cilia. They are both required not only to build the cilium but to maintain it and drive transport along this structure. Our work has reached a point where we consider it hugely beneficial to combine our efforts to better understand how these complexes work, not just in isolation, but in the context of the other large multi-protein machines that are required for cilia function. The dynein-2 and kinesin-2 microtubule motors provides a fundamental link between microtubule motor function, protein trafficking, and cilia function because of its function in driving intraflagellar transport (IFT) within cilia. The Stephens lab was the first to define the subunit composition of the dynein-2 motor in humans. The Nakayama lab have made major advances in our understanding of how kinesin-2 integrates with the IFT machinery. This ambitious project seeks to combine our expertise in protein interaction analysis and cell imaging to define how dynein-2 and kinesin-2 interact with, and work in concert with, the other major ciliary machines called IFT-A, IFT-B and the BBSome. We aim to provide a complete picture of the molecular interactions of the dynein-2 complex with other critical components of the system, the kinesin-2 motor, the BBSome and the IFT particles, IFT-A and IFT-B, using a combination of molecular cell biology approaches including advanced microscopy and proteomics. Our current BBSRC-funded work has developed proteomics approaches that have identified key interacting proteins that seem to direct the assembly and function of dynein-2. This is a frontier bioscience project that seeks to understand fundamental processes in cell biology. That said, the formation of cilia, tight control of cilia-based signalling pathways, and the control of entry to and exit from the cell cycle are fundamental to normal health as well as having potential long-term impact on human and animal health. Ciliary signals include those that control early human development as well as others that occur throughout life to control metabolism. Key pharmaceuticals targeting common cancers are also directed against ciliary signalling pathway. A full understanding of the structure and function of cilia is key to a diverse array of fields and has relevance from the earliest stages of human development and throughout life.

初级纤毛从几乎所有人类细胞的表面伸出，作为信号平台。它们对人类和动物的发育是必不可少的，在整个生命过程中也需要控制与骨骼、肾脏功能、大脑中的信号传递和更多身体功能相关的通路。这些纤毛是由一系列大型多蛋白质复合体构建和维持的。我们在英国和日本的两个实验室为我们了解所涉及的关键机器做出了重大贡献。最近，我们各自定义了纤毛中两个微管运动蛋白复合体--动力蛋白-2和动力蛋白-2的功能。它们不仅需要建造纤毛，而且还需要维护纤毛和推动沿这个结构的运输。我们的工作已经到了这样一个地步，我们认为联合努力更好地了解这些复合体是如何工作的，这是非常有益的，不仅是在孤立的情况下，而且在纤毛功能所需的其他大型多蛋白质机器的背景下也是如此。动力蛋白-2和动蛋白-2微管马达在驱动纤毛内鞭毛运输(IFT)方面发挥着重要作用，在微管运动功能、蛋白质转运和纤毛功能之间起着重要的作用。斯蒂芬斯实验室是第一个确定人类动力蛋白-2马达亚基组成的实验室。Nakayama实验室在我们理解Kinesin-2如何与IFT机器整合方面取得了重大进展。这个雄心勃勃的项目寻求结合我们在蛋白质相互作用分析和细胞成像方面的专业知识，以确定dynein-2和kinesin-2如何与称为IFT-A、IFT-B和BBSome的其他主要纤毛机器相互作用和协同工作。我们的目标是利用包括高级显微镜和蛋白质组学在内的分子细胞生物学方法，提供Dynein-2复合体与系统的其他关键组件，Kinesin-2马达、BBSome和IFT颗粒，IFT-A和IFT-B的分子相互作用的完整图景。我们目前由BBSRC资助的工作已经开发出蛋白质组学方法，已经确定了关键的相互作用蛋白，似乎指导了Dynein-2的组装和功能。这是一个前沿性的生物科学项目，旨在了解细胞生物学的基本过程。也就是说，纤毛的形成、纤毛信号通路的严格控制以及细胞周期的进出控制是正常健康的基础，并对人和动物的健康具有潜在的长期影响。纤毛信号包括那些控制人类早期发育的信号，以及其他在整个生命中发生的控制新陈代谢的信号。针对常见癌症的关键药物也针对纤毛信号通路。充分了解纤毛的结构和功能是一系列不同领域的关键，从人类发展的最早阶段到整个生命都具有相关性。

项目成果

Roles for CEP170 in cilia function and dynein-2 assembly.

CEP170 在纤毛功能和动力蛋白 2 组装中的作用。

• DOI: 10.1101/2023.11.20.567836
• 发表时间: 2023
• 作者: Weijman J