Tsao Che-Chia, Gorovsky Martin A
Department of Biology, University of Rochester, Rochester, NY 14627, USA.
Mol Biol Cell. 2008 Apr;19(4):1450-61. doi: 10.1091/mbc.e07-05-0403. Epub 2008 Jan 16.
Intraflagellar transport (IFT) particles are multiprotein complexes that move bidirectionally along the cilium/flagellum. The Tetrahymena IFT172 gene encodes a protein with an N-terminal WD domain (WDD) and a C-terminal repeat domain (RPD). Epitope-tagged Ift172p localized to the basal body and in cilia along the axoneme, and IFT172 knockout cells lost cilia and motility. Using serial deletion constructs to rescue the knockout cells, we found that neither the WDD nor the RPD alone is sufficient to assemble cilia. Ift172p containing only the WDD or the RPD failed to enter cilia. Constructs with a partial truncation of the RPD still rescued although cilia were assembled less efficiently, indicating that the WDD and a part of the RPD are sufficient for anterograde transport. Partial truncation of the RPD caused the accumulation of truncated Ift172p itself and of Ift88p at ciliary tips, suggesting that IFT turnaround or retrograde transport was affected. These results implicate different regions of Ift172p in different steps of the IFT process.
鞭毛内运输(IFT)颗粒是沿纤毛/鞭毛双向移动的多蛋白复合物。嗜热四膜虫IFT172基因编码一种具有N端WD结构域(WDD)和C端重复结构域(RPD)的蛋白质。表位标记的Ift172p定位于基体以及沿着轴丝的纤毛中,并且IFT172基因敲除细胞失去了纤毛和运动能力。使用系列缺失构建体来拯救敲除细胞,我们发现单独的WDD或RPD都不足以组装纤毛。仅包含WDD或RPD的Ift172p无法进入纤毛。RPD部分截短的构建体仍然能够拯救细胞,尽管纤毛组装效率较低,这表明WDD和部分RPD足以进行顺行运输。RPD的部分截短导致截短的Ift172p自身以及Ift88p在纤毛顶端积累,这表明IFT周转或逆行运输受到了影响。这些结果表明Ift172p的不同区域在IFT过程的不同步骤中发挥作用。