Modification of Escherichia coli membranes in the prereplicative phase of phage T4 infection. Specificity of association and quantitation of bound phage proteins.
Reinfection of Escherichia coli with the bacterial virus T4 causes modifications of the properties of the host cell envelope during the preeplicative phase of the lytic cycle. These changes include altered densities cell enveloped and their subfractions, morphological modifications of membrane vesicles, and association of newly synthesized proteins with the host cell envelope. Polypeptide analysis by high resolution electrophoresis on polyacrylamide slab gels in dodecyl sulfate revealed that most of some 30 prereplicative phage-coded polypeptides are attached to this structure. Different means of cell disruption and selective extraction procedures, such as variations of ionic strength, removal of divalent cations, and the addition of chaotropic agents or detergents were used to study the characteristics of these attachments. Many proteins appeared to be artifactually absorbed or weakly bound to the envelope, Separation of cell walls from plasma membranes showed that all of the tightly bound proteins were associated withthe cell membrane fraction. The partitioning of phage proteins between the different fractions was monitored using 12 polypeptides which were identified as products of distinct phage genes. Of these, 8 were eliminated as potential membrane markers. Four polypeptides, the products of genes rIIA, rIIB, 39, and 52 were operationally defined as membrane proteins. The number of molecules of the 12 identified phage gene products, synthesized during a single lytic cycle, was determined. The results allowed the estimation of the concentration in the membrane of those proteins which were found to be quantitatively associated with that structure. Association of phage proteins with the cell envelope was found to be unaffected by mutations in any of the identified phage polypeptides.
D000596:Amino Acids / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism*; D003090:Coliphages / Q000378:metabolism*; D004267:DNA Viruses / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism* / Q000648:ultrastructure; D005796:Genes; D006146:Guanidines; D008274:Magnesium / Q000494:pharmacology; D009154:Mutation; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D013104:Spheroplasts / Q000378:metabolism; D013997:Time Factors; D014764:Viral Proteins / Q000096:biosynthesis*
D000596:Amino Acids; D006146:Guanidines; D010446:Peptide Fragments; D014764:Viral Proteins; D004492:Edetic Acid; D008274:Magnesium