The traffic ATPase PilF interacts with the inner membrane platform of the DNA translocator and type IV pili from Thermus thermophilus
The traffic ATPase PilF interacts with the inner membrane platform of the DNA translocator and type IV pili from Thermus thermophilus
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交通 ATP 酶 PilF 与来自嗜热栖热菌的 DNA 易位器和 IV 型菌毛的内膜平台相互作用
DOI：
10.1002/2211-5463.12548
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发表时间：
2019
期刊：
影响因子：
2.6
通讯作者：
Averhoff
中科院分区：
文献类型：
--
作者：
Salzer;Averhoff
细菌适应不断变化的环境的一个主要驱动力是通过自然转化从环境中摄取裸露的DNA，这使得获得新的能力成为可能。高分子量DNA的摄取由跨越整个细胞外周的复杂运输机制介导。这种DNA转运蛋白催化双链DNA的结合和分裂，以及单链DNA转运到细胞质中，在细胞质中与染色体重组。嗜热菌Thermus thermophilus具有最高的转化频率，是分析这种大分子转运机制的结构和功能的模型系统。运输活性由运输ATP酶PilF提供动力，PilF是一种形成六聚体复合物的可溶性蛋白。在这里，我们证明了PilF通过ATP结合蛋白PilM与DNA转运蛋白的内膜组装平台（包括PilMNO）物理结合。与PilMNO或PilMN的结合刺激PilF的ATP酶活性~ 2倍，而当单独与PilM或PilN结合时没有刺激。ATP结合缺陷的PilMK 26 A变体仍然结合PilF，并与PilN一起刺激PilF介导的ATP酶活性。PilF的独特之处在于在其N末端具有三个保守的GSPII（一般分泌途径II）结构域（A-C）。缺失分析显示，GSPII结构域中没有一个是结合PilMN所必需的，但GSPIIC是PilF对PilMN介导的ATP水解的刺激所必需的。我们的数据表明，PilM是一种偶联蛋白，其通过PilMNO组装平台在物理和功能上将可溶性马达ATP酶PilF连接到DNA转运子。
A major driving force for the adaptation of bacteria to changing environments is the uptake of naked DNA from the environment by natural transformation, which allows the acquisition of new capabilities. Uptake of the high molecular weight DNA is mediated by a complex transport machinery that spans the entire cell periphery. This DNA translocator catalyzes the binding and splitting of double‐stranded DNA and translocation of single‐stranded DNA into the cytoplasm, where it is recombined with the chromosome. The thermophilic bacteriumThermus thermophilusexhibits the highest transformation frequencies reported and is a model system to analyze the structure and function of this macromolecular transport machinery. Transport activity is powered by the traffic ATPase PilF, a soluble protein that forms hexameric complexes. Here, we demonstrate that PilF physically binds to an inner membrane assembly platform of the DNA translocator, comprising PilMNO, via the ATP‐binding protein PilM. Binding to PilMNO or PilMN stimulates the ATPase activity of PilF ~ 2‐fold, whereas there is no stimulation when binding to PilM or PilN alone. A PilMK26Avariant defective in ATP binding still binds PilF and, together with PilN, stimulates PilF‐mediated ATPase activity. PilF is unique in having three conserved GSPII (general secretory pathway II) domains (A–C) at its N terminus. Deletion analyses revealed that none of the GSPII domains is essential for binding PilMN, but GSPIIC is essential for PilMN‐mediated stimulation of ATP hydrolysis by PilF. Our data suggest that PilM is a coupling protein that physically and functionally connects the soluble motor ATPase PilF to the DNA translocator via the PilMNO assembly platform.
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