Elsevier

Virology

Volume 171, Issue 1, July 1989, Pages 229-238
Virology

Quantitation of the adsorption and penetration stages of bacteriophage φ6 infection

https://doi.org/10.1016/0042-6822(89)90530-8Get rights and content

Abstract

The enveloped dsRNA bacteriophage φ6 uses the pilus of Pseudomonas syringae as its receptor. It enters the host cell by fusion of the virus envelope with the host outer membrane, followed by penetration of the cytoplasmic membrane by the phage nucleocapsid. In this investigation we quantitated the adsorption and penetration of φ6wt and a host range mutant, φ6h1s, to five bacterial strains. Adsorption rate constants were measured for the different phage-host combinations, the constant for φ6wt with the standard host was 3.3 × 10−10 ml/min. Infections with 14C-labeled phage at different phage/cell ratios were used to measure the numbers of adsorbing and entering virions/sensitive cell. At high phage/cell ratios (200–250) the standard host adsorbed on the average 35–40 wild-type virions/cell, the saturation level being somewhat higher. It was shown that at phage/host cell ratios of 0.1–1 practically every virion produces an infectious center. The average number of entering phage particles per infectious center reached saturation around the phage/cell ratio of 50 and did not exceed 3 for the standard host. The φ6 preparations used in this study had a specific infectivity of 0.7–0.9.

References (44)

  • J.F. Lehman et al.

    Isolation of new mutants of bacteriophage φ6

    Virology

    (1979)
  • L. Mindich et al.

    Genetic studies of temperature-sensitive and nonsense mutants of bacteriophage φ6

    Virology

    (1976)
  • L. Mindich et al.

    The morphogenesis of bacteriophage φ6: Particles formed by nonsense mutants

    Virology

    (1976)
  • H.R. Revel et al.

    Generation of cDNA clones of the bacteriophage φ6 segmented dsRNA genome: Characterization and expression of L segment clones

    Virology

    (1986)
  • A. Rimon et al.

    Transcription and replication of bacteriophage φ6 RNA

    Virology

    (1978)
  • A. Rimon et al.

    Temperature-sensitive mutants of bacteriophage φ6 defective in both transcription and replication

    Virology

    (1978)
  • J.F. Sinclair et al.

    RNA synthesis during infection with bacteriophage φ6

    Virology

    (1976)
  • S.J. Usala et al.

    Displacement of parental RNA strands during in vitro transcription by bacteriophage φ6 nucleocapsids

    Cell

    (1980)
  • M.H. Adams

    Bacteriophages

  • D.H. Bamford et al.

    Ultrastructure and life cycle of the lipid-containing bacteriophage φ6

    J. Gen. Virol.

    (1976)
  • D.H. Bamford et al.

    Membrane fusion in prokaryotes: Bacteriophage φ6 membrane fuses with the Pseudomonas syringae outer membrane

    EMBO J.

    (1987)
  • D.A. Cuppels et al.

    In vitro translation of the three bacteriophage φ6 RNAs

    J. Virol.

    (1980)
  • Cited by (130)

    • Inline-tandem purification of viruses from cell lysate by agarose-based chromatography

      2022, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
    • The Molecular Mechanism of Cellular Attachment for an Archaeal Virus

      2019, Structure
      Citation Excerpt :

      Absorbtion rate constants for other viruses presented in Table S1 were obtained from the following references: SMV1 (Uldahl et al., 2016), SSV9 (Bautista et al., 2015), and SIRV2 (Quemin et al., 2013). Bacteriophages: T1-T4 (Puck et al., 1951), ϕ6 (Olkkonen and Bamford, 1989), and PV22 (Zhilenkov et al., 2006). Halophilic archaeal viruses SH1, HHTV1 His1, and His2 (Svirskaite et al., 2016).

    • Asymmetrical flow field-flow fractionation in purification of an enveloped bacteriophage ϕ6

      2018, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
      Citation Excerpt :

      Fractions (1 mL) were collected starting from the beginning of the elution gradient. Proteins were analysed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) using 15% (w/v) acrylamide in the separation gel [19] and Coomassie staining. Nucleic acids were visualized by ethidium bromide staining of the stacking gel.

    • Vesicle-like virion of Haloarcula hispanica pleomorphic virus 3 preserves high infectivity in saturated salt

      2016, Virology
      Citation Excerpt :

      In order to test HHPV3 sensitivity to these negative stains, 1× purified viruses were incubated in the undiluted stain solution (1:1) for the appropriate staining times (see above) at 22 °C, after which infectivity was determined by plaque assay. Virion components were analyzed using tricine-sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE; 16% acrylamide in the separation gel) (Olkkonen and Bamford, 1989). Gels were stained with Coomassie Brilliant Blue R 250 (Serva) for proteins and Sudan Black B (Sigma Aldrich) for lipids.

    • Asymmetric flow field flow fractionation methods for virus purification

      2016, Journal of Chromatography A
      Citation Excerpt :

      The precipitated proteins were collected by centrifugation (Eppendorf centrifuge 5415D, 13,000 rpm, 30 min, 4 °C) and resuspended in 1.5 × SDS-PAGE sample buffer [42]. Boiled samples were analyzed in SDS polyacrylamide (SDS-PAGE) gels made in-house that used 16% acrylamide in the separation gels [42]. Proteins were visualized with Coomassie stain.

    View all citing articles on Scopus
    View full text