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

总结 纤毛是重要的细胞器，其功能包括从细胞间信号传递到体内平衡的产生 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”，因为它们共有纤毛结构或功能缺陷的病因学。尽管在世界上有着明确的角色， 中枢神经系统、四肢、中轴骨骼、肾脏、气道、大脑和生殖系统的发育 道，我们的理解的机制，支配纤毛发生和纤毛介导的发育 因为纤毛需要数百种不同的蛋白质， 生物起源和功能，通过包含未知的多种蛋白质的广泛相互作用网络起作用。 功能我们建议在这里研究几个大的多蛋白质组装体是必要的适当的纤毛 以确定这些复合物在纤毛发生的关键步骤中的作用，包括募集 的蛋白质的基体，鞭毛内运输和招聘特定的鞭毛内货物。这 格兰特结合了定向机械实验，蛋白质组学，3D建模，体内细胞生物学和测试， 在模式生物中的人类疾病等位基因，以了解关键纤毛蛋白及其 相互作用伴侣影响适当的纤毛形成，以及这些基因中的特定突变如何导致出生 缺陷通过关注在发育和疾病中表现出重要性的蛋白质， 的作用机制是已知的，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的新细胞过程的理解。反过来， 这些发现应该为一系列先天性疾病提供更深入的了解， 口-面-指综合征与完全致命的短肋多指畸形

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.