Architecture of the type IVa pilus machine.

Architecture of the type IVa pilus machine.
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DOI:
10.1126/science.aad2001
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发表时间:
2016-03-11
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Jensen GJ
Jensen GJ
中科院分区:
其他
文献类型:
--
作者:
Chang YW;Rettberg LA;Treuner-Lange A;Iwasa J;Søgaard-Andersen L;Jensen GJ

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IVa型菌毛是在许多细菌中观察到的丝状细胞表面结构。它们通过伸展、附着在表面,然后收缩来拉动细胞向前。我们使用完整的黄色黏液球菌细胞的低温电子断层扫描,以3到4纳米的分辨率,在原位观察IVa型菌毛和组装和收缩它们的蛋白质机器(IVa型菌毛机器,或T4PM),包括有毛和无毛的状态。我们发现,T4PM包括一个外膜孔、四周质和细胞质中四个相互连接的环状结构、一个细胞质圆盘和穹窿以及一个周围质茎。通过对缺乏定义的T4PM蛋白或单个蛋白与标签融合的突变体进行系统成像,我们绘制了所有10个T4PM核心成分和次要支柱的位置,从而提供了对菌毛组装、结构和功能的见解。细菌型IVa菌毛机(T4PM)。左边描绘了两个t4pm,跨越革兰氏阴性细菌细胞的包膜。t4pm伸展和收缩毛以拉动细胞向前。本文提供的结构数据支持这样的假设,即细胞质中的ATP水解导致内膜中的一种转换蛋白旋转,促进了毛蛋白亚基从内膜转移到生长的毛上。在收缩过程中,这个过程是相反的。
Type IVa pili are filamentous cell surface structures observed in many bacteria. They pull cells forward by extending, adhering to surfaces, and then retracting. We used cryo–electron tomography of intact Myxococcus xanthus cells to visualize type IVa pili and the protein machine that assembles and retracts them (the type IVa pilus machine, or T4PM) in situ, in both the piliated and nonpiliated states, at a resolution of 3 to 4 nanometers. We found that T4PM comprises an outer membrane pore, four interconnected ring structures in the periplasm and cytoplasm, a cytoplasmic disc and dome, and a periplasmic stem. By systematically imaging mutants lacking defined T4PM proteins or with individual proteins fused to tags, we mapped the locations of all 10 T4PM core components and the minor pilins, thereby providing insights into pilus assembly, structure, and function. Bacterial type IVa pilus machine (T4PM). Two T4PMs are depicted on the left, spanning the envelope of a Gram-negative bacterial cell. T4PMs extend and retract pili to pull cells forward. The structural data presented here support the hypothesis that ATP hydrolysis in the cytoplasm causes an adapter protein in the inner membrane to rotate, facilitating the transfer of pilin subunits from the inner membrane onto the growing pilus. The process is reversed during retraction.
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