Functional genomic screen for modulators of ciliogenesis and cilium length.
Functional genomic screen for modulators of ciliogenesis and cilium length.
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作者：
初级纤毛是进化上保守的细胞器，组织不同的信号通路。初级纤毛的形成或功能缺陷与一系列人类疾病和发育异常有关。模型生物体的遗传筛选发现了纤毛组装和维护的核心机制。然而，协调初级纤毛生物发生与其他细胞过程（包括细胞骨架组织、囊泡运输和细胞间粘附）的调节分子仍有待鉴定。在这里，我们报告了使用 RNA 干扰 (RNAi) 来鉴定参与纤毛发生控制的人类基因的功能基因组筛选的结果。筛选鉴定出 36 个正纤毛发生调节剂和 13 个负纤毛发生调节剂，其中包括参与肌动蛋白动力学和囊泡运输的分子。进一步的研究表明，阻断肌动蛋白组装通过稳定中心体周围前纤毛室（PPC）来促进纤毛发生，PPC是一种先前未表征的紧凑的囊小管结构，在纤毛发生的早期阶段储存用于纤毛的跨膜蛋白。 PPC 通过回收内体标记物进行标记。此外，参与内吞再循环途径的调节剂的敲低影响了 PPC 的形成以及纤毛发生。我们的结果揭示了将肌动蛋白动力学和内吞循环与纤毛发生结合起来的关键调控步骤，并为未来的研究提供了潜在的靶分子。
Primary cilia are evolutionarily conserved cellular organelles that organize diverse signaling pathways. Defects in the formation or function of primary cilia are associated with a spectrum of human diseases and developmental abnormalities. Genetic screens in model organisms have discovered core machineries of cilium assembly and maintenance. However, regulatory molecules that coordinate the biogenesis of primary cilia with other cellular processes, including cytoskeletal organization, vesicle trafficking and cell-cell adhesion, remain to be identified. Here we report the results of a functional genomic screen using RNA interference (RNAi) to identify human genes involved in ciliogenesis control. The screen identified 36 positive and 13 negative ciliogenesis modulators, which include molecules involved in actin dynamics and vesicle trafficking. Further investigation demonstrated that blocking actin assembly facilitates ciliogenesis by stabilizing the pericentrosomal preciliary compartment (PPC), a previously uncharacterized compact vesiculotubular structure storing transmembrane proteins destined for cilia during the early phase of ciliogenesis. PPC was labeled by recycling endosome markers. Moreover, knockdown of modulators that are involved in the endocytic recycling pathway affected the formation of PPC as well as ciliogenesis. Our results uncover a critical regulatory step that couples actin dynamics and endocytic recycling with ciliogenesis, and also provide potential target molecules for future study.
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DOI：
10.1152/ajprenal.00273.2001
发表时间：
2002-03-01
影响因子：
4.2
作者：
Yoder, BK;Tousson, A;Balkovetz, DF
通讯作者：
Balkovetz, DF
影响因子：
4.8
作者：
Valentin, E;Ghomashchi, F;Lambeau, G
通讯作者：
Lambeau, G
影响因子：
64.5
作者：
Gerdes JM;Davis EE;Katsanis N
通讯作者：
Katsanis N
影响因子：
64.8
作者：
Corbit, KC;Aanstad, P;Reiter, JF
通讯作者：
Reiter, JF
影响因子：
64.5
作者：
Nachury, Maxence V.;Loktev, Alexander V.;Jackson, Peter K.
通讯作者：
Jackson, Peter K.