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低温电子显微镜揭示了 Dot/Icm T4SS 中的新种特异性蛋白和对称元素。

Cryo-EM reveals new species-specific proteins and symmetry elements in the Dot/Icm T4SS.

机构信息

Department of Pharmacology, University of Minnesota, Minneapolis, United States.

Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, United States.

出版信息

Elife. 2021 Sep 14;10:e70427. doi: 10.7554/eLife.70427.

DOI:10.7554/eLife.70427
PMID:34519271
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8486379/
Abstract

is an opportunistic pathogen that causes the potentially fatal pneumonia known as Legionnaires' disease. The pathology associated with infection depends on bacterial delivery of effector proteins into the host via the membrane spanning Dot/Icm type IV secretion system (T4SS). We have determined sub-3.0 Å resolution maps of the Dot/Icm T4SS core complex by single particle cryo-EM. The high-resolution structural analysis has allowed us to identify proteins encoded outside the Dot/Icm genetic locus that contribute to the core T4SS structure. We can also now define two distinct areas of symmetry mismatch, one that connects the C18 periplasmic ring (PR) and the C13 outer membrane cap (OMC) and one that connects the C13 OMC with a 16-fold symmetric dome. Unexpectedly, the connection between the PR and OMC is DotH, with five copies sandwiched between the OMC and PR to accommodate the symmetry mismatch. Finally, we observe multiple conformations in the reconstructions that indicate flexibility within the structure.

摘要

是一种机会致病菌，可引起称为军团病的潜在致命性肺炎。与感染相关的病理学取决于细菌通过跨膜 Dot/Icm 型 IV 型分泌系统 (T4SS) 将效应蛋白递送到宿主中。我们通过单颗粒冷冻电镜确定了 Dot/Icm T4SS 核心复合物的亚 3.0 Å 分辨率图谱。高分辨率结构分析使我们能够识别出编码在 Dot/Icm 遗传基因座之外的有助于核心 T4SS 结构的蛋白质。我们现在还可以定义两个不同的对称不匹配区域，一个连接 C18 周质环 (PR) 和 C13 外膜帽 (OMC)，另一个连接 C13 OMC 和 16 倍对称圆顶。出乎意料的是，PR 和 OMC 之间的连接是 DotH，五个拷贝夹在 OMC 和 PR 之间以适应对称不匹配。最后，我们在重建中观察到多个构象，表明结构内的灵活性。

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