Wingfield Jenna L, Mengoni Ilaria, Bomberger Heather, Jiang Yu-Yang, Walsh Jonathon D, Brown Jason M, Picariello Tyler, Cochran Deborah A, Zhu Bing, Pan Junmin, Eggenschwiler Jonathan, Gaertig Jacek, Witman George B, Kner Peter, Lechtreck Karl
Department of Cellular Biology, University of Georgia, Athens, United States.
College of Engineering, University of Georgia, Athens, United States.
Elife. 2017 May 31;6:e26609. doi: 10.7554/eLife.26609.
Intraflagellar transport (IFT) trains, multimegadalton assemblies of IFT proteins and motors, traffic proteins in cilia. To study how trains assemble, we employed fluorescence protein-tagged IFT proteins in . IFT-A and motor proteins are recruited from the cell body to the basal body pool, assembled into trains, move through the cilium, and disperse back into the cell body. In contrast to this 'open' system, IFT-B proteins from retrograde trains reenter the pool and a portion is reused directly in anterograde trains indicating a 'semi-open' system. Similar IFT systems were also observed in and IMCD3 cells. FRAP analysis indicated that IFT proteins and motors of a given train are sequentially recruited to the basal bodies. IFT dynein and tubulin cargoes are loaded briefly before the trains depart. We conclude that the pool contains IFT trains in multiple stages of assembly queuing for successive release into the cilium upon completion.
鞭毛内运输（IFT）列车是由IFT蛋白和马达组成的多兆道尔顿组装体，负责在纤毛中运输蛋白质。为了研究列车如何组装，我们在……中使用了荧光蛋白标记的IFT蛋白。IFT-A和马达蛋白从细胞体被招募到基体池，组装成列车，穿过纤毛，然后分散回到细胞体。与这种“开放”系统不同，逆行列车中的IFT-B蛋白重新进入池，并且一部分直接在顺行列车中被重新利用，这表明是一种“半开放”系统。在……和IMCD3细胞中也观察到了类似的IFT系统。荧光恢复后光漂白（FRAP）分析表明，给定列车的IFT蛋白和马达是依次被招募到基体的。IFT动力蛋白和微管蛋白货物在列车出发前被短暂装载。我们得出结论，该池中包含处于多个组装阶段的IFT列车，排队等待完成后依次释放到纤毛中。
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