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Published July 2017 | Supplemental Material + Submitted
Journal Article Open

In vivo structures of an intact type VI secretion system revealed by electron cryotomography

Abstract

The type VI secretion system (T6SS) is a versatile molecular weapon used by many bacteria against eukaryotic hosts or prokaryotic competitors. It consists of a cytoplasmic bacteriophage tail‐like structure anchored in the bacterial cell envelope via a cytoplasmic baseplate and a periplasmic membrane complex. Rapid contraction of the sheath in the bacteriophage tail‐like structure propels an inner tube/spike complex through the target cell envelope to deliver effectors. While structures of purified contracted sheath and purified membrane complex have been solved, because sheaths contract upon cell lysis and purification, no structure is available for the extended sheath. Structural information about the baseplate is also lacking. Here, we use electron cryotomography to directly visualize intact T6SS structures inside Myxococcus xanthus cells. Using sub‐tomogram averaging, we resolve the structure of the extended sheath and membrane‐associated components including the baseplate. Moreover, we identify novel extracellular bacteriophage tail fiber‐like antennae. These results provide new structural insights into how the extended sheath prevents premature disassembly and how this sophisticated machine may recognize targets.

Additional Information

© 2017 The Authors. Received 13 February 2017; Revised 16 March 2017; Accepted 5 April 2017; Published online 9 May 2017. Data availability: The sub‐tomogram averages of the M. xanthus T6SS that support the findings of this study have been deposited in the Electron Microscopy Data Bank with accession codes EMD‐8600 (membrane‐associated region), EMD‐8601 (extended sheath), EMD‐8602 (contracted sheath). The coordinates of the extended and contracted sheath models have been deposited in the Protein Data Bank with accession code 5URW and 5URX, respectively. We thank Prof. Hong Z. Zhou and Dr. Peng Ge for providing the initial concept of how to place the MyTssBC model in the sub‐tomogram average of extended T6SS sheath and Dr. Catherine Oikonomou for discussions and editorial assistance. This work was supported by NIH grant R01 AI127401 to G.J.J. Author contributions: Y‐WC and LAR collected, processed, and analyzed the electron cryotomography data. Y‐WC built the models. DRO performed the bioinformatics analysis. Y‐WC, DRO and GJJ wrote the paper. The authors declare that they have no conflict of interest.

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Supplemental Material - inline-supplementary-material-1.pdf

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August 21, 2023
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