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

Ravi K. Lokareddy, Chun-Feng David Hou, Francesca Forti, Stephano M. Iglesias, Fenglin Li, Mikhail Pavlenok, David S. Horner, Michael Niederweis, Federica Briani () and Gino Cingolani ()
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Ravi K. Lokareddy: 1825 University Blvd
Chun-Feng David Hou: The State University of New Jersey
Francesca Forti: Università degli Studi di Milano
Stephano M. Iglesias: Perelman School of Medicine at the University of Pennsylvania
Fenglin Li: Perelman School of Medicine at the University of Pennsylvania
Mikhail Pavlenok: 845 19th Street South
David S. Horner: Università degli Studi di Milano
Michael Niederweis: 845 19th Street South
Federica Briani: Università degli Studi di Milano
Gino Cingolani: 1825 University Blvd

Nature Communications, 2024, vol. 15, issue 1, 1-20

Abstract: 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 3398 amino acid virion-associated RNA polymerase (vRNAP). 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 but dispensable for infecting mutants with a truncated lipopolysaccharide devoid of the O-antigen. We determine that DEV vRNAP is part of a three-gene operon conserved in 191 Schitoviridae genomes. We propose these three proteins are ejected into the host to form a genome ejection motor spanning the cell envelope. We posit that the design principles of the DEV ejection apparatus are conserved in all Schitoviridae.

Date: 2024
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DOI: 10.1038/s41467-024-52752-1

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