The tail sheath of bacteriophage N4 interacts with the Escherichia coli receptor

J Bacteriol. 2009 Jan;191(2):525-32. doi: 10.1128/JB.01423-08. Epub 2008 Nov 14.

Abstract

Unlike other characterized phages, the lytic coliphage N4 must inject the 360-kDa virion RNA polymerase (vRNAP), in addition to its 72-kbp genome, into the host for successful infection. The process of adsorption to the host sets up and elicits the necessary conformational changes in the virion to allow genome and vRNAP injection. Infection of suppressor and nonsuppressor strains, Escherichia coli W3350 supF and E. coli W3350, with a mutant N4 isolate (N4am229) harboring an amber mutation in Orf65 yielded virions containing (N4gp65(+)) and lacking (N4gp65(-)) gp65, respectively. N4gp65(+) but not N4gp65(-) phage was able to adsorb to the host. Recombinant gp65 with a hexahistidine tag at the N terminus or hexahistidine and c-myc tags at the C terminus was able to complement N4gp65(-) virions in vivo and in vitro. Immunogold detection of gp65 in vivo complemented virions revealed its localization at the N4 tail. Finally, we show both in vitro and in vivo that gp65 interacts with the previously determined N4 outer membrane receptor, NfrA.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / metabolism*
  • Bacteriophage N4 / genetics
  • Bacteriophage N4 / metabolism*
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli / virology*
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism*
  • Glycoproteins / genetics
  • Glycoproteins / metabolism*
  • Mutation
  • Protein Binding
  • Receptors, Virus / genetics
  • Receptors, Virus / metabolism*
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / metabolism*
  • Virion / genetics
  • Virion / metabolism

Substances

  • Bacterial Outer Membrane Proteins
  • Escherichia coli Proteins
  • Glycoproteins
  • NfrA protein, E coli
  • Receptors, Virus
  • Viral Envelope Proteins