Roles of linker region flanked by transmembrane and peptidoglycan binding region of PomB in energy conversion of the Vibrio flagellar motor

IF 1.3 4区 生物学 Q4 CELL BIOLOGY Genes to Cells Pub Date : 2024-02-14 DOI:10.1111/gtc.13102
Yusuke Miyamura, Tatsuro Nishikino, Hiroaki Koiwa, Michio Homma, Seiji Kojima
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Abstract

The flagellar components of Vibrio spp., PomA and PomB, form a complex that transduces sodium ion and contributes to rotate flagella. The transmembrane protein PomB is attached to the basal body T-ring by its periplasmic region and has a plug segment following the transmembrane helix to prevent ion flux. Previously we showed that PomB deleted from E41 to R120 (Δ41–120) was functionally comparable to the full-length PomB. In this study, three deletions after the plug region, PomB (Δ61–120), PomB (Δ61–140), and PomB (Δ71–150), were generated. PomB (Δ61–120) conferred motility, whereas the other two mutants showed almost no motility in soft agar plate; however, we observed some swimming cells with speed comparable for the wild-type cells. When the two PomB mutants were introduced into a wild-type strain, the swimming ability was not affected by the mutant PomBs. Then, we purified the mutant PomAB complexes to confirm the stator formation. When plug mutations were introduced into the PomB mutants, the reduced motility by the deletion was rescued, suggesting that the stator was activated. Our results indicate that the deletions prevent the stator activation and the linker and plug regions, from E41 to S150, are not essential for the motor function of PomB but are important for its regulation.

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PomB 跨膜和肽聚糖结合区两侧的连接区在弧菌鞭毛运动的能量转换中的作用。
弧菌的鞭毛成分 PomA 和 PomB 形成一个复合体,可转导钠离子并促进鞭毛旋转。跨膜蛋白 PomB 通过其外质区连接到基体 T 环上,并在跨膜螺旋之后有一个堵塞段,以防止离子流动。此前我们曾发现,从 E41 到 R120(Δ41-120)缺失的 PomB 在功能上与全长 PomB 相当。在这项研究中,我们生成了插接区之后的三个缺失区,即 PomB(Δ61-120)、PomB(Δ61-140)和 PomB(Δ71-150)。PomB(Δ61-120)具有运动能力,而其他两个突变体在软琼脂平板上几乎没有运动能力;不过,我们观察到一些游动细胞的速度与野生型细胞相当。将这两个 PomB 突变体导入野生型菌株后,突变 PomB 并未影响其游动能力。然后,我们纯化了突变型 PomAB 复合物,以确认定子的形成。当将插接突变引入 PomB 突变体时,缺失导致的运动能力下降得到了挽救,这表明定子被激活了。我们的研究结果表明,这些缺失阻止了定子的激活,从E41到S150的连接区和插接区对PomB的运动功能并不重要,但对其调控却很重要。
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来源期刊
Genes to Cells
Genes to Cells 生物-细胞生物学
CiteScore
3.40
自引率
0.00%
发文量
71
审稿时长
3 months
期刊介绍: Genes to Cells provides an international forum for the publication of papers describing important aspects of molecular and cellular biology. The journal aims to present papers that provide conceptual advance in the relevant field. Particular emphasis will be placed on work aimed at understanding the basic mechanisms underlying biological events.
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