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Structure paper

TitleIntegrative structural analysis of Pseudomonas phage DEV reveals a genome ejection motor.
Journal, issue, pagesNat Commun, Vol. 15, Issue 1, Page 8482, Year 2024
Publish dateOct 1, 2024
AuthorsRavi K Lokareddy / Chun-Feng David Hou / Francesca Forti / Stephano M Iglesias / Fenglin Li / Mikhail Pavlenok / David S Horner / Michael Niederweis / Federica Briani / Gino Cingolani /
PubMed AbstractDEV 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 ...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.
External linksNat Commun / PubMed:39353939 / PubMed Central
MethodsEM (single particle)
Resolution3.1 - 4.7 Å
Structure data

43629

8vxq

EMDB-43629, PDB-8vxq:
Cryo-EM structure of phage DEV ejection proteins gp72:gp73
Method: EM (single particle) / Resolution: 3.1 Å

44517

9bgm

EMDB-44517, PDB-9bgm:
Pseudomonas phage DEV neck and tail (portal, head-to-tail and tail tube proteins)
Method: EM (single particle) / Resolution: 3.1 Å

44518

9bgn

EMDB-44518, PDB-9bgn:
Pseudomonas phage DEV 5-fold vertex (major coat protein)
Method: EM (single particle) / Resolution: 3.3 Å

44519

9bgo

EMDB-44519, PDB-9bgo:
Pseudomonas phage DEV gp72 ejection protein (pre-ejection conformation)
Method: EM (single particle) / Resolution: 4.2 Å

45776

9cod

EMDB-45776, PDB-9cod:
C15 symmetrized DEV collar
Method: EM (single particle) / Resolution: 4.7 Å

Source
  • pseudomonas phage vb_paep_dev (virus)
  • pseudomonas aeruginosa (bacteria)
KeywordsVIRAL PROTEIN / phage / bacteriophage / STRUCTURAL PROTEIN / outer membrane protein / gp73 / gp74 / DEV / portal / tail hub / tail tube / complex / gp75 / gp83 / gp80 / virion coat / gp77 / virion scaffolding / gp72 / ejection protein / gp53

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