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PreprintCited by 15Bacterial Genetics and BiotechnologyEscherichia coli research studiesBiochemical and Structural Characterization

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Structure of a Rhs effector clade domain identifies new mechanistic insights into type VI secretion system toxin delivery.

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McGowan, Sheena, Hayes, Brooke, Harper, Marina, Venugopal, Hariprasad, Lewis, Jessica, Wright, Amy, Steele, Joel, Steer, David, Schittenhelm, Ralf, Boyce, John·2023·Nature Communications
DOI 10.21203/rs.3.rs-3632996/v1PMC PMC11461821

Preprint — published as 10.1038/s41467-024-52950-x · Paper page

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Abstract

The type VI secretion system (T6SS) is a molecular machine utilised by many Gram-negative bacteria to deliver antibacterial toxins directly into adjacent bacteria, often providing a competitive advantage. Rearrangement hotspot (Rhs) effectors are known T6SS cargo effectors, but the precise delivery and activation of these toxins is poorly defined. We present the structure of a novel T6SS Rhs effector (Tse15) from the multidrug resistant nosocomial pathogen Acinetobacter baumannii. Tse15 forms a triple layered β-cocoon Rhs domain with an N-terminal α-helical clade domain and an unfolded C-terminal toxin domain located entirely inside the Rhs cage. We identify that Tse15 is cleaved into three domains, through two independent auto-cleavage events involving aspartyl protease activity for toxin self-cleavage and a nucleophilic glutamic acid for the cleavage of the N-terminal clade domain. Proteomic analyses showed that the N-terminal clade and toxin domains, but not the β-strand rich Rhs cage, are delivered outside of the cell, suggesting a novel mechanism for Rhs toxin delivery and activation. Our findings suggest that this delivery mechanism requires an interaction between the N-terminal clade and toxin domains, with the clade domain acting as the internal chaperone to mediate tethering of the toxin to the T6SS machinery. Conservation of the clade domain in other Gram-negative bacteria suggest this may be a common mechanism for T6SS toxin delivery.

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Metadata

Authors
McGowan, S., Hayes, B., Harper, M., Venugopal, H., Lewis, J., Wright, A., Steele, J., Steer, D., Schittenhelm, R., & Boyce, J.
Affiliations
Monash University (AU)
Corresponding author
  • McGowan, Sheena
  • Boyce, John
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2023-12-13
Volume
15
Issue
1
Pages
8709-8709
ISSN
2041-1723 (ISSN-L)
DOI
10.21203/rs.3.rs-3632996/v1
PMC
PMC11461821
OpenAlex
W4403204445
Cited by
15
References
1
Citations by year
2026: 9 · 2025: 4 · 2024: 2
FWCI
3.96
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • Australian Research Council (LE120100090, LE200100045)
  • National Health and Medical Research Council (1165036, 1128981)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Immunology and Microbiology › Microbiology › Escherichia coli research studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
Keywords
type VI secretion system, Acinetobacter baumannii, toxin delivery, toxin secretion, Gram-negative bacteria, protein structure
MeSH terms
Type VI Secretion Systems, Protein Domains, Amino Acid Sequence, Bacterial Proteins, Bacterial Toxins, Models, Molecular, Crystallography, X-Ray, Proteomics, Acinetobacter baumannii

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