Rab35 controls cilium length, function and membrane composition
Rab35 controls cilium length, function and membrane composition
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DOI：
10.15252/embr.201847625
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发表时间：
2019-08-21
期刊：
影响因子：
7.7
通讯作者：
Blacque, Oliver E.
中科院分区：
文献类型：
--
作者：
Kuhns, Stefanie;Seixas, Cecilia;Blacque, Oliver E.
Rab 和 Arl 鸟嘌呤核苷酸结合 (G) 蛋白调节初级纤毛形成、功能和组成所必需的运输途径，初级纤毛是与 Sonic Hedgehog (Shh) 信号传导和纤毛病相关的感觉装置。在这里，我们利用哺乳动物细胞和斑马鱼，揭示了 Rab35 的纤毛功能，Rab35 是一种具有内吞循环、肌动蛋白重塑和胞质分裂作用的多任务 G 蛋白。通过 siRNA、吗啉代或敲除导致的 Rab35 缺失会缩短哺乳动物细胞和斑马鱼左右组织体（库普弗囊泡）中的纤毛长度，并导致与活动纤毛相关的左右不对称缺陷。与这些观察结果一致，GFP-Rab35 定位于纤毛，GEF (DENND1B) 和 GAP (TBC1D10A) Rab35 调节因子也是如此，它们也调节纤毛长度和 Rab35 纤毛定位。哺乳动物 Rab35 还控制 Shh 信号调节因子的纤毛膜水平，促进 Smoothened 的纤毛靶向，限制 Arl13b 和肌醇多磷酸 5-磷酸酶 (INPP5E) 的纤毛积累。 Rab35 还调节纤毛 PI(4,5)P-2 水平并与 Arl13b 相互作用。总之，我们的研究结果证明了 Rab35 在调节纤毛长度、功能和膜组成中的作用，并表明 Rab35 参与了控制 Shh 信号调节器纤毛水平的途径。
Rab and Arl guanine nucleotide-binding (G) proteins regulate trafficking pathways essential for the formation, function and composition of primary cilia, which are sensory devices associated with Sonic hedgehog (Shh) signalling and ciliopathies. Here, using mammalian cells and zebrafish, we uncover ciliary functions for Rab35, a multitasking G protein with endocytic recycling, actin remodelling and cytokinesis roles. Rab35 loss via siRNAs, morpholinos or knockout reduces cilium length in mammalian cells and the zebrafish left-right organiser (Kupffer's vesicle) and causes motile cilia-associated left-right asymmetry defects. Consistent with these observations, GFP-Rab35 localises to cilia, as do GEF (DENND1B) and GAP (TBC1D10A) Rab35 regulators, which also regulate ciliary length and Rab35 ciliary localisation. Mammalian Rab35 also controls the ciliary membrane levels of Shh signalling regulators, promoting ciliary targeting of Smoothened, limiting ciliary accumulation of Arl13b and the inositol polyphosphate 5-phosphatase (INPP5E). Rab35 additionally regulates ciliary PI(4,5)P-2 levels and interacts with Arl13b. Together, our findings demonstrate roles for Rab35 in regulating cilium length, function and membrane composition and implicate Rab35 in pathways controlling the ciliary levels of Shh signal regulators.