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PreprintCited by 18Bacteriophages and microbial interactionsAdvanced Electron Microscopy Techniques and ApplicationsBacterial Genetics and Biotechnology

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Integrative structural analysis of Pseudomonas phage DEV reveals a genome ejection motor

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Cingolani, Gino, Lokareddy, Ravi, Hou, Chun-Feng, Forti, Francesca, Iglesias, Stephano, Li, Fenglin, Pavlenok, Mikhail, Niederweis, Michael, Briani, Federica·2024·Nature Communications
DOI 10.21203/rs.3.rs-3941185/v1PMC PMC11445570

Preprint — published as 10.1038/s41467-024-52752-1 · Paper page

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Abstract

DEV is an obligatory lytic Pseudomonas phage of the N4-like genus, recently reclassified as Schitoviridae. The DEV genome encodes 91 ORFs, including a 3,398 amino acid virion-associated RNA polymerase. Here, we describe the complete architecture of DEV, determined using a combination of cryo-electron microscopy localized reconstruction, biochemical methods, and genetic knockouts. We built de novo structures of all capsid factors and tail components involved in host attachment. We demonstrate that DEV long tail fibers are essential for infection of Pseudomonas aeruginosa and dispensable for infecting mutants with a truncated lipopolysaccharide devoid of the O-antigen. We identified DEV ejection proteins and, unexpectedly, found that the giant DEV RNA polymerase, the hallmark of the Schitoviridae family, is an ejection protein. We propose that DEV ejection proteins form a genome ejection motor across the host cell envelope and that these structural principles are conserved in all Schitoviridae.

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Metadata

Authors
Cingolani, G., Lokareddy, R., Hou, C.-F., Forti, F., Iglesias, S., Li, F., Pavlenok, M., Niederweis, M., & Briani, F.
Affiliations
  • University of Alabama at Birmingham (US)
  • Rutgers, The State University of New Jersey (US)
  • University of Milan (IT)
  • University of Pennsylvania (US)
Corresponding author
  • Cingolani, Gino
  • Briani, Federica
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2024-02-21
Volume
15
Issue
1
Pages
8482-8482
ISSN
2041-1723 (ISSN-L)
DOI
10.21203/rs.3.rs-3941185/v1
PMC
PMC11445570
OpenAlex
W4403039848
Cited by
18
References
1
Citations by year
2026: 5 · 2025: 10 · 2024: 3
FWCI
4.94
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by-nc-nd)
  • Research Square (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • Archivio Istituzionale della Ricerca (Universita Degli Studi Di Milano) (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
  • U.S. Department of Health and Human Services (R35GM140733, S10 OD030457, R01GM100888)
  • Università degli Studi di Milano
  • Papua New Guinea University of Technology
  • Fondazione per la Ricerca sulla Fibrosi Cistica
  • National Institutes of Health (75N91019D00024, 1s10od030457-01a1)
  • Centers for Disease Control and Prevention (S10 OD030457)
  • National Cancer Institute (75N91019D00024)
  • National Institute of General Medical Sciences (U24GM129541, R01GM100888, R35GM140733)
  • SLAC National Accelerator Laboratory
  • Common Fund
Topics
  • Life Sciences › Immunology and Microbiology › Virology › Bacteriophages and microbial interactions
  • 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
Keywords
phage genome structure, host attachment, lipopolysaccharide O-antigen, Pseudomonas aeruginosa, capsid structure, tail structure, operon

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