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PreprintCited by 37Advanced Electron Microscopy Techniques and ApplicationsBacterial Genetics and BiotechnologyBiochemical and Structural Characterization

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CryoEM structure of the Vibrio cholerae Type IV competence pilus secretin PilQ

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Weaver, Sara J., Sazinsky, Matthew H., Dalia, Triana N., Dalia, Ankur B., Jensen, Grant J.·2020·Nature Communications
DOI 10.1101/2020.03.03.975896PMC PMC7545093

Preprint — published as 10.1038/s41467-020-18866-y · Paper page

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Abstract

Natural transformation is the process by which bacteria take up genetic material from their environment and integrate it into their genome by homologous recombination. It represents one mode of horizontal gene transfer and contributes to the spread of traits like antibiotic resistance. In Vibrio cholerae , the Type IV competence pilus is thought to facilitate natural transformation by extending from the cell surface, binding to exogenous DNA, and retracting to thread this DNA through the outer membrane secretin, PilQ. A lack of structural information has hindered our understanding of this process, however. Here, we solved the first ever high-resolution structure of a Type IV competence pilus secretin. A functional tagged allele of VcPilQ purified from native V. cholerae cells was used to determine the cryoEM structure of the PilQ secretin in amphipol to ∼2.7 Å. This structure highlights for the first time key differences in the architecture of the Type IV competence pilus secretin from the Type II and Type III Secretin System secretins. Based on our cryoEM structure, we designed a series of mutants to interrogate the mechanism of PilQ. These experiments provide insight into the channel that DNA likely traverses to promote the spread of antibiotic resistance via horizontal gene transfer by natural transformation. We prove that it is possible to reduce pilus biogenesis and natural transformation by sealing the gate, suggesting VcPilQ as a new drug target.

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Metadata

Authors
Weaver, S. J., Sazinsky, M. H., Dalia, T. N., Dalia, A. B., & Jensen, G. J.
Affiliations
  • California Institute of Technology (US)
  • Pomona College (US)
  • Indiana University Bloomington (US)
Corresponding author
Jensen, Grant J.
ORCID
Venue
Nature Communications
Publisher
openRxiv
Published
2020-03-04
Volume
11
Issue
1
Pages
5080-5080
ISSN
2041-1723 (ISSN-L)
DOI
10.1101/2020.03.03.975896
PMC
PMC7545093
OpenAlex
W3092186467
Cited by
37
References
1
Citations by year
2026: 7 · 2025: 6 · 2024: 5 · 2023: 7 · 2022: 6 · 2021: 4 · 2020: 2
FWCI
1.94
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • CaltechAUTHORS (California Institute of Technology) (submittedVersion, cc-by-nc-nd)
  • CaltechAUTHORS (California Institute of Technology) (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Howard Hughes Medical Institute
  • U.S. Department of Health and Human Services (R35GM128674, AI127401)
  • California Institute of Technology
  • National Institutes of Health
  • National Institute of General Medical Sciences (R35GM128674)
  • National Institute of Allergy and Infectious Diseases (R01AI127401)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
Keywords
Vibrio cholerae, natural competence, natural transformation, horizontal gene transfer, DNA uptake, antibiotic resistance spread, druggable targets, Proteobacteria
MeSH terms
Protein Domains, Cysteine, Membrane Proteins, Models, Molecular, Mutation, Phylogeny, Fimbriae, Bacterial, Secretin, Transformation, Bacterial, Vibrio cholerae, Cryoelectron Microscopy, Bacterial Secretion Systems

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