Chaperone mediated coupling of subunit availability to activation of flagellar Type III Secretion

Chaperone mediated coupling of subunit availability to activation of flagellar Type III Secretion
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分子伴侣介导亚基可用性与鞭毛 III 型分泌激活的耦合

DOI:
10.1101/2020.01.10.902387
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发表时间:
2020
期刊:
--
影响因子:
--
通讯作者:
Bryant O
Bryant O
中科院分区:
--
文献类型:
--
作者:
Bryant O

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细菌鞭毛亚基通过鞭毛 III 型分泌系统 (fT3SS) 穿过细胞膜输出,该系统由质子动力 (pmf) 和特殊的 ATP 酶提供动力,使鞭毛输出门能够利用 pmf 电势 (ΔΨ)。输出门激活是由 ATP 酶茎 FliJ 介导的,但如何调节该过程以防止在没有亚基货物的情况下浪费性耗散 pmf 尚不清楚。在这里,我们表明 FliJ 输出门的激活受到鞭毛输出伴侣的调节。 FliJ 结合未负载的分子伴侣,并且通过使用对 FliJ 结合具有特异性缺陷的新型分子伴侣变体,我们发现这种相互作用的破坏会减弱运动性和同源亚基输出。我们在体外证明了伴侣分子和 FlhA 输出门组件竞争结合 FliJ,并在体内证明了当亚基水平较低时细胞中存在的未负载伴侣分子会隔离 FliJ 以防止输出门激活并减弱亚基输出。我们的数据表明了一种机制,伴侣分子将亚基货物的可用性与 fT3SS 的 pmf 驱动的输出耦合起来。
Bacterial flagellar subunits are exported across the cell membrane by the flagellar Type III Secretion System (fT3SS), powered by the proton motive force (pmf) and a specialized ATPase that enables the flagellar export gate to utilize the pmf electric potential (ΔΨ). Export gate activation is mediated by the ATPase stalk, FliJ, but how this process is regulated to prevent wasteful dissipation of pmf in the absence of subunit cargo is not known. Here, we show that FliJ activation of the export gate is regulated by flagellar export chaperones. FliJ binds unladen chaperones and, by using novel chaperone variants specifically defective for FliJ binding, we show that disruption of this interaction attenuates motility and cognate subunit export. We demonstrate in vitro that chaperones and the FlhA export gate component compete for binding to FliJ, and show in vivo that unladen chaperones, which would be present in the cell when subunit levels are low, sequester FliJ to prevent activation of the export gate and attenuate subunit export. Our data indicate a mechanism whereby chaperones couple availability of subunit cargo to pmf‐driven export by the fT3SS.
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