Mapping the ciliary interactome, an extensive protein interaction network underlying human ciliopathies

摘要 纤毛是必需的细胞器，其功能从细胞间信号传递到产生内环境平衡。 管状器官中的液体流动。因此，一系列人类先天性疾病被描述为 “纤毛病症”，因为它们都有纤毛结构或功能缺陷的病因。尽管有明确的角色在 中枢神经系统、四肢、中轴骨、肾脏、呼吸道、大脑和生殖的发育 纤维束，我们对纤毛发生和纤毛介导的发育机制的理解 模式仍然不完整，尤其是因为正常的纤毛需要数百种不同的蛋白质 生物发生和功能，通过包含各种未知蛋白质的广泛相互作用网络发挥作用 功能。我们建议在这里研究几个大型多蛋白组合，它们是正常纤毛所必需的。 形成，以确定这些复合体在纤毛发生的关键步骤中的作用，包括招募 蛋白质的运输、鞭毛内的运输和特定鞭毛内货物的募集。这 格兰特结合了定向机械实验、蛋白质组学、3D建模、体内细胞生物学和 人类疾病等位基因在模式生物中的作用，以了解关键纤毛蛋白及其 相互作用的伙伴影响适当的纤毛形成，以及这些基因的特定突变如何导致出生。 缺陷。通过关注在发育和疾病中证明重要的蛋白质，但对这些蛋白质没有 作用机制尚不清楚，这里提出的实验将提供重要的新的广度和深度 我们对纤毛介导的发育模式和纤毛生物学中新的细胞过程的理解。反过来, 这些发现应该会为一系列先天性疾病提供更多的洞察力，从相对轻微的 口腔-面部-指端综合征致全致死性短肋骨多指畸形。

SUMMARY Cilia are essential organelles, with functions ranging from cell-cell signaling to the generation of homeostatic fluid flow in tubular organs. Consequently, an array of human congenital diseases has been characterized as “ciliopathies,” because they share an etiology of defective cilia structure or function. Despite clear roles in the development of the central nervous system, limbs, axial skeleton, kidneys, airway, brain, and reproductive tracts, our understanding of the mechanisms that govern ciliogenesis and cilia-mediated developmental patterning remain incomplete, not least because hundreds of different proteins are required for proper cilia biogenesis and function, acting via an extensive interaction network containing diverse proteins of unknown function. We propose here to study several large multi-protein assemblies that are essential for proper cilia formation in order to determine the roles of these complexes in key steps in ciliogenesis, including recruitment of proteins to the basal body, intraflagellar transport and recruitment of specific intraflagellar cargoes. This grant combines directed mechanistic experiments, proteomics, 3D modeling, in vivo cell biology, and testing of human disease alleles in model organisms to understand mechanisms by which key ciliary proteins and their interaction partners effect proper cilia formation, and how specific mutations in these genes lead to birth defects. By focusing on proteins with demonstrated importance in development and disease, but for which no mechanism of action is yet known, experiments proposed here will provide important new breadth and depth to our understanding cilia-mediated developmental patterning and novel cell processes in ciliary biology. In turn, these findings should provide greater insight to a range of congenital diseases ranging from the relatively mild Oral-Facial-Digital syndrome to the wholly lethal Short Rib Polydactyly.