Functional dissection of structural regions of the Thermus thermophilus competence protein PilW: Implication in secretin complex stability, natural transformation and pilus functions.
Functional dissection of structural regions of the Thermus thermophilus competence protein PilW: Implication in secretin complex stability, natural transformation and pilus functions.
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嗜热栖热菌能力蛋白 PilW 结构区域的功能剖析：对促胰液素复合物稳定性、自然转化和菌毛功能的影响
DOI：
10.1016/j.bbamem.2021.183666
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Averhoff
中科院分区：
文献类型：
--
作者：
Averhoff
DNA从环境中摄取到细菌细胞质是由一种大分子运输机制介导的，这种机制在结构和功能上类似于IV型菌毛(T4P)，确实，DNA转运体和T4P具有共同的成分。一种是分泌素PilQ，它被组装成均聚复合体，形成高度动态的外膜(OM)通道，介导菌毛挤出和DNA摄取。PilQ如何与发动机相互作用仍然是个谜。在这里，我们使用生化和遗传学技术来研究PilQ和PilW的相互作用，PilW是一种独特的蛋白质，对自然转化和T4P排泄是必不可少的。嗜热菌。PilQ和PilW在有机质中形成高分子质量的复合体。嗜热菌。当P ilW缺失时，不再观察到PilQ复合体，而只观察到PilQ单体，伴随着DNA摄取的丧失以及T4P和抽动运动的丧失。一个Δ-PilT2/Δ-PilW双突变体的发现表明，PilW对于PilQ复合体的稳定组装是重要的。为了分析PilW的不同区域的作用，我们构建了部分缺失(PilW∆2-40，PilW∆50-150，PilW∆163-216和PilW∆216-292)，并研究了它们对DNA摄取、PilQ复合体形成和T4P功能(如抽动运动、生物膜形成和细胞-细胞相互作用)的影响。这些研究表明，菌毛动力学需要在PilW中有一个中心无序区域。我们认为PilW是连接转运体ATPase的蛋白质网络的一部分，以驱动DNA转运体和T4P的不同功能。
Uptake of DNA from the environment into the bacterial cytoplasm is mediated by a macromolecular transport machinery that is similar in structure and function to type IV pili (T4P) and, indeed, DNA translocator and T4P share common components. One is the secretin PilQ which is assembled into homopolymeric complexes forming highly dynamic outer membrane (OM) channels mediating pilus extrusion and DNA uptake. How PilQ interacts with the motor is still enigmatic. Here, we have used biochemical and genetic techniques to study the interaction of PilQ with PilW, a unique protein which is essential for natural transformation and T4P extrusion ofT. thermophilus. PilQ and PilW form high molecular mass complexes in the OM ofT. thermophilus. WhenpilWwas deleted, PilQ complexes were no longer observed but only PilQ monomers, accompanied by a loss of DNA uptake as well as a loss of T4P and twitching motility. Piliation of a ΔpilT2/ΔpilWdouble mutant suggests that PilW is important for stable assembly of PilQ complexes. To analyze the role of different regions of PilW, partial deletions (pilW∆2–40, pilW∆50–150, pilW∆163–216andpilW∆216–292) were generated and the effect on DNA uptake, PilQ complex formation and T4P functions such as twitching motility, biofilm formation and cell-cell interaction was studied. These studies revealed that a central disordered region in PilW is required for pilus dynamics. We propose that PilW is part of a protein network that connects the transport ATPase to drive different functions of the DNA translocator and T4P.