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通过珠粒测定法研究铜绿假单胞菌鞭毛马达中两个定子系统的动态。

Dynamics of the Two Stator Systems in the Flagellar Motor of Pseudomonas aeruginosa Studied by a Bead Assay.

机构信息

Hefei National Laboratory for Physical Sciences at the Microscale and Department of Physics, University of Science and Technology of Chinagrid.59053.3a, Hefei, Anhui, China.

出版信息

Appl Environ Microbiol. 2021 Nov 10;87(23):e0167421. doi: 10.1128/AEM.01674-21. Epub 2021 Sep 15.

Abstract

We developed a robust bead assay for studying flagellar motor behavior of Pseudomonas aeruginosa. Using this assay, we studied the dynamics of the two stator systems in the flagellar motor. We found that the two sets of stators function differently, with MotAB stators providing higher total torque and MotCD stators ensuring more stable motor speed. The motors in wild-type cells adjust the stator compositions according to the environment, resulting in an optimal performance in environmental exploration compared to that of mutants with one set of stators. The bead assay we developed in this investigation can be further used to study P. aeruginosa chemotaxis at the level of a single cell using the motor behavior as the chemotaxis output. Cells of Pseudomonas aeruginosa possess a single polar flagellum, driven by a rotatory motor powered by two sets of torque-generating units (stators). We developed a robust bead assay for studying the behavior of the flagellar motor in P. aeruginosa, by attaching a microsphere to shortened flagellar filament and using it as an indicator of motor rotation. Using this assay, we revealed the dynamics of the two stator systems in the flagellar motor and found that the motors in wild-type cells adjust the stator compositions according to the environment, resulting in an optimal performance in environmental exploration compared to that of mutants with one set of stators.

摘要

我们开发了一种稳健的珠子检测法来研究铜绿假单胞菌的鞭毛运动行为。使用这种检测方法,我们研究了鞭毛运动中的两个定子系统的动力学。我们发现这两套定子的功能不同,MotAB 定子提供更高的总扭矩,而 MotCD 定子确保更稳定的电机速度。与只有一套定子的突变体相比,野生型细胞中的电机根据环境调整定子组成,从而在环境探索方面表现出最佳性能。我们在这项研究中开发的珠子检测法可以进一步用于使用电机行为作为趋化性输出来研究铜绿假单胞菌的单细胞趋化性。铜绿假单胞菌具有单个极鞭毛,由由两组产生扭矩的单元(定子)驱动的旋转式电机驱动。我们开发了一种稳健的珠子检测法来研究铜绿假单胞菌鞭毛运动中的电机行为,通过将微球附着在缩短的鞭毛丝上,并将其用作电机旋转的指示。使用这种检测方法,我们揭示了鞭毛运动中两个定子系统的动力学,发现野生型细胞中的电机根据环境调整定子组成,从而在环境探索方面表现出最佳性能,与只有一套定子的突变体相比。

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Nat Microbiol. 2020 Dec;5(12):1553-1564. doi: 10.1038/s41564-020-0788-8. Epub 2020 Sep 14.
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