Kunitomo Hirofumi, Iino Yuichi
Molecular Genetics Research Laboratory, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
Genes Cells. 2008 Jan;13(1):13-25. doi: 10.1111/j.1365-2443.2007.01147.x.
Cilia and flagella play critical roles in cell motility, development and sensory perception in animals. Formation and maintenance of cilia require a conserved protein transport system called intraflagellar transport (IFT). Here, we show that Caenorhabditis elegans dyf-11 encodes an evolutionarily conserved protein required for cilium biogenesis. dyf-11 is expressed in most of the ciliated neurons and is regulated by DAF-19, a crucial transcription factor for ciliary genes in C. elegans. dyf-11 mutants exhibit stunted cilia, fluorescent dye-filling defects (Dyf) of sensory neurons, and abnormal chemotaxis (Che). Cell- and stage-specific rescue experiments indicated that DYF-11 is required for formation and maintenance of sensory cilia in cell-autonomous manner. Fluorescent protein-tagged DYF-11 localizes to cilia and moves antero- and retrogradely via IFT. Analysis of DYF-11 movement in bbs mutants further suggested that DYF-11 is likely associated with IFT complex B. Domain analysis using DYF-11 deletion constructs revealed that the coiled-coil region is required for proper localization and ciliogenesis. We further show that Traf3ip1/MIP-T3, the mammalian orthologue of DYF-11, localizes to cilia in the MDCK renal epithelial cells.
纤毛和鞭毛在动物的细胞运动、发育及感官感知中发挥着关键作用。纤毛的形成与维持需要一种名为鞭毛内运输(IFT)的保守蛋白运输系统。在此,我们表明秀丽隐杆线虫的dyf-11编码一种纤毛生物发生所需的进化保守蛋白。dyf-11在大多数具纤毛的神经元中表达,并受DAF-19调控,DAF-19是秀丽隐杆线虫中纤毛基因的关键转录因子。dyf-11突变体表现出纤毛发育不良、感觉神经元的荧光染料填充缺陷(Dyf)以及异常趋化性(Che)。细胞和阶段特异性拯救实验表明,DYF-11以细胞自主方式参与感觉纤毛的形成与维持。荧光蛋白标记的DYF-11定位于纤毛,并通过IFT进行顺行和逆行移动。对bbs突变体中DYF-11移动的分析进一步表明,DYF-11可能与IFT复合物B相关。使用DYF-11缺失构建体进行的结构域分析表明,卷曲螺旋区域对于正确定位和纤毛发生是必需的。我们进一步表明,DYF-11的哺乳动物同源物Traf3ip1/MIP-T3定位于MDCK肾上皮细胞的纤毛中。