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Requirement of IFT-B–BBSome complex interaction in export of GPR161 from cilia

机译：纤毛中GPR161出口中IFT-B-BBS的复杂相互作用的要求

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The intraflagellar transport (IFT) machinery, which includes the IFT-A and IFT-B complexes, mediates bidirectional trafficking of ciliary proteins. In addition to these complexes, the BBSome, which is composed of eight subunits that are encoded by the causative genes of Bardet-Biedl syndrome (BBS), has been proposed to connect the IFT machinery to ciliary membrane proteins, such as G protein-coupled receptors, to mediate their export from cilia. However, little is known about the connection between the IFT machinery and the BBSome. Using the visible immunoprecipitation assay, we here identified the interaction between IFT38 from the IFT-B complex and BBS1, BBS2 and BBS9 from the BBSome. Furthermore, by analyzing phenotypes ofIFT38-knockout cells exogenously expressing wild-type IFT38 or its mutant lacking the ability to interact with BBS1+BBS2+BBS9, we showed that knockout cells expressing the IFT38 mutant have restored ciliogenesis; however, similar toBBS1-knockout cells, they demonstrated significant accumulation of GPR161 within cilia upon stimulation of Hedgehog signaling. These results indicate that the IFT-B–BBSome interaction is required for the export of GPR161 across the ciliary gate.
机译：鞭毛内运输（IFT）机制，包括IFT-A和IFT-B复合物，介导睫状蛋白的双向运输。除了这些复合物外，还提出了由八种亚基组成的BBSome，这些亚基由Bardet-Biedl综合征（BBS）的致病基因编码，可以将IFT机制连接到睫状膜蛋白上，例如G蛋白偶联受体，介导纤毛的出口。但是，对于IFT机械和BBSome之间的连接了解甚少。使用可见的免疫沉淀测定法，我们在这里鉴定了IFT-B复合物的IFT38与BBSome的BBS1，BBS2和BBS9之间的相互作用。此外，通过分析外源表达野生型IFT38或其缺乏与BBS1 + BBS2 + BBS9相互作用能力的突变体的IFT38敲除细胞的表型，我们表明表达IFT38突变体的敲除细胞已恢复纤毛形成。然而，类似于BBS1敲除细胞，它们在刺激刺猬信号后显示出纤毛内GPR161的大量积累。这些结果表明，IFR-B-BBSome相互作用是跨睫状口出口GPR161所必需的。

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