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A Highlights from MBoC Selection: Tetrahymena RIB72A and RIB72B are microtubule inner proteins in the ciliary doublet microtubules

机译:MBoC选择集锦:四膜虫RIB72A和RIB72B是睫状双联微管中的微管内部蛋白

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摘要

Doublet and triplet microtubules are essential and highly stable core structures of centrioles, basal bodies, cilia, and flagella. In contrast to dynamic cytoplasmic micro­tubules, their luminal surface is coated with regularly arranged microtubule inner proteins (MIPs). However, the protein composition and biological function(s) of MIPs remain poorly understood. Using genetic, biochemical, and imaging techniques, we identified Tetrahymena RIB72A and RIB72B proteins as ciliary MIPs. Fluorescence imaging of tagged RIB72A and RIB72B showed that both proteins colocalize to Tetrahymena cilia and basal bodies but assemble independently. Cryoelectron tomography of RIB72A and/or RIB72B knockout strains revealed major structural defects in the ciliary A-tubule involving MIP1, MIP4, and MIP6 structures. The defects of individual mutants were complementary in the double mutant. All mutants had reduced swimming speed and ciliary beat frequencies, and high-speed video imaging revealed abnormal highly curved cilia during power stroke. Our results show that RIB72A and RIB72B are crucial for the structural assembly of ciliary A-tubule MIPs and are important for proper ciliary motility.
机译:双线和三线微管是至关重要的,中心粒,基体,纤毛和鞭毛的核心结构高度稳定。与动态细胞质微管相反,它们的腔表面被规则排列的微管内部蛋白(MIP)覆盖。但是,人们对MIP的蛋白质组成和生物学功能知之甚少。使用遗传,生化和成像技术,我们确定了四膜虫RIB72A和RIB72B蛋白为睫状MIP。标记的RIB72A和RIB72B的荧光成像显示,这两种蛋白共定位于纤毛膜纤毛和基体,但独立组装。 RIB72A和/或RIB72B敲除菌株的低温电子层析成像显示,睫状A管中的主要结构缺陷涉及MIP1,MIP4和MIP6结构。单个突变体的缺陷在双突变体中是互补的。所有突变体的游泳速度和纤毛跳动频率均降低,并且高速视频成像显示在中风期间异常弯曲的纤毛异常。我们的结果表明,RIB72A和RIB72B对于睫状A管MIP的结构组装至关重要,并且对于适当的睫状活动性也很重要。

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