Centrosome regulation of trafficking, ciliogenesis and cilia-dependent signaling

项目摘要/摘要 中心体是细胞的中心组织中心，为有丝分裂的纺锤体和 纤毛。各种人类疾病从缺陷到中心体，因为它们同时影响有丝分裂和纤毛 功能。我们发现唐氏综合症患者的细胞既有中心体缺陷，又有缺陷 纤毛。这为理解唐氏综合征的细胞生物学基础开辟了一条重要的新途径。 由于初级纤毛的丧失在发育上是致命的，我们的研究将探索细微变化对 中心体在控制纤毛的形成和功能方面起作用。21三体纤毛功能减退的中心 细胞，我们发现编码在21号染色体上的Pericentin基因的表达上调是 必须且充分地抑制纤毛形成和功能所需的组件的移动 从细胞质到中心体和纤毛。周丝蛋白是一种中心体和胞内转运蛋白 支架蛋白在细胞周期中急剧增加和减少，纤毛时水平最低 形成发生在有丝分裂期间最高。我们认为，对周毛菌素的精细控制可以确保纤毛 构成和功能。然而，我们不明白为什么周围素蛋白适度增加1.5倍 发现唐氏综合症对纤毛有如此深刻的影响。我们将测试如何减少和增加 周中心素水平影响中心体、间期微管景观以及卫星和囊泡。 贩运，以及纤毛形成和信号传递的结果是什么。优秀的培训 皮尔逊实验室的本科生、研究生和博士后研究人员都有机会。 我们将继续使用和发展我们严格的培训和实验程序，以与NIH保持一致 提高严密性和重复性的指导方针。与其他电子专业实验室的合作 显微镜和机器学习定量分析将扩大我们研究的创新和影响 细胞内交易。总而言之，这项提案将阐明如何精确控制人类疾病 周丝蛋白与细胞周期相协调，以确保适当的初级纤毛形成和信号传递。 正常发育所必需的。这项应用旨在阐明基本的细胞生物学事件在 人类常见缺陷的细胞内运输、纤毛形成和纤毛依赖信号 疾病。

PROJECT SUMMARY / ABSTRACT Centrosomes are the central organizing centers of cells, performing essential functions for mitotic spindles and cilia. Diverse human diseases arise from defects to centrosomes because they impact both mitotic and ciliary functions. We discovered that cells from individuals with Down syndrome have both defective centrosomes and cilia. This opens an important new avenue for understanding the cell biological basis of Down syndrome. Because primary cilia loss is developmentally lethal, our studies will explore the role that subtle changes to centrosomes have in controlling cilia formation and function. Central to the reduced cilia function in Trisomy 21 cells, we discovered that elevated expression of the Pericentrin gene, encoded on chromosome 21, is necessary and sufficient to repress the movement of components required for both cilia formation and function from the cytoplasm to the centrosome and cilium. Pericentrin is a centrosome and intracellular trafficking scaffold protein that dramatically increases and decreases during the cell cycle, with levels lowest when cilia formation occurs and highest during mitosis. We propose that exquisite control of Pericentrin ensures cilia formation and function. However, we do not understand why the modest 1.5-fold increase in Pericentrin protein found in Down syndrome has such a profound impact on cilia. We will test how decreased and increased Pericentrin levels impact centrosomes, the interphase microtubule landscape, and satellite and vesicular trafficking, and what the consequent outcomes for cilia formation and signaling are. Excellent training opportunities exist for undergraduate and graduate students and postdoctoral researchers in the Pearson lab. We will continue to use and develop our rigorous training and experimental routines in alignment with the NIH guidelines for increasing rigor and reproducibility. Collaborations with other labs that specialize in electron microscopy and machine learning quantitative analyses will expand the innovation and impact of our studies of intracellular trafficking. In summary, this proposal will illuminate how the precise control of the human disease protein, Pericentrin, is coordinated with the cell cycle to ensure proper primary cilia formation and signaling required for normal development. This application aims to elucidate fundamental cell biological events in intracellular trafficking, cilia formation and cilia-dependent signaling that are commonly defective in human disease.