Genetics for Pseudoalteromonas provides tools to manipulate marine bacterial virus PM2

Hanna M. Kivelä, Stefania Madonna, Mart Krupovìč, M. Luisa Tutino, Jaana K H Bamford

Research output: Contribution to journalArticle

Abstract

The genetic manipulation of marine double-stranded DNA (dsDNA) bacteriophage PM2 (Corticoviridae) has been limited so far. The isolation of an autonomously replicating DNA element of Pseudoalteromonas haloplanktis TAC125 and construction of a shuttle vector replicating in both Escherichia coli and Pseudoalteromonas enabled us to design a set of conjugative shuttle plasmids encoding tRNA suppressors for amber mutations. Using a host strain carrying a suppressor plasmid allows the introduction and analysis of nonsense mutations in PM2. Here, we describe the isolation and characterization of a suppressor-sensitive PM2 sus2 mutant deficient in the structural protein P10. To infect and replicate, PM2 delivers its 10-kbp genome across the cell envelopes of two gram-negative Pseudoalteromonas species. The events leading to the internalization of the circular supercoiled dsDNA are puzzling. In a poorly understood process that follows receptor recognition, the virion capsid disassembles and the internal membrane fuses with the host outer membrane. While beginning to unravel the mechanism of this process, we found that protein P10 plays an essential role in the host cell penetration.

Original languageEnglish
Pages (from-to)1298-1307
Number of pages10
JournalJournal of Bacteriology
Volume190
Issue number4
DOIs
Publication statusPublished - Feb 2008

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

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    Kivelä, H. M., Madonna, S., Krupovìč, M., Tutino, M. L., & Bamford, J. K. H. (2008). Genetics for Pseudoalteromonas provides tools to manipulate marine bacterial virus PM2. Journal of Bacteriology, 190(4), 1298-1307. https://doi.org/10.1128/JB.01639-07