Abstract
When Escherichia coli B, labeled by prior growth in (14)C-glucose, are infected with T4 phage there is a rapid release of (14)C-nondialyzable material into the medium. About half of this material is derived from the cell envelope as evidenced by its content of phospholipid and lipopolysaccharide and its buoyant density upon isopycnic ultracentrifugation of 1.19 g/cm(3). It is similar in its gross chemical and physical properties to envelope material released at a lower rate from growing uninfected cells or from cells whose protein synthesis is inhibited by chloramphenicol (22). The rate of release of this envelope material at a multiplicity of infection (MOI) of 10 is greatest in the first minute after infection, and release is completed by 4 min. The rate of its release, as a function of MOI at 2 min after infection, is greatest at low MOI (e.g., MOI 2 and 4); in addition, the release does not continue above MOI 30. The main conclusion derived from the data is that phage, as part of the process of adsorption and injection of DNA, cause an increased release of envelope substance from the cells. With the assumption that all of the envelope material released is derived from the outer envelope, it is estimated that uninfected cells release 20 to 30% of their outer envelope per hour, whereas infected cells release 30% in 2 min at MOI 30. Further, because release does not continue at high MOI, this phenomenon is not considered to be a direct cause of lysis from without. Data are also presented on the amounts of other non-dialyzable (14)C-components released and on the differences in the kinetics of release from chloramphenicol-treated cells compared to phage-infected cells. To avoid the possibility that the release is due to phage lysozyme which is an adventitious "contaminant" of wild-type phage, a phage mutant (T4BeG59s) devoid of this enzyme was used in these experiments.
MeSH Terms
Adsorption
Carbon Radioisotopes
Cell Wall/metabolism
Centrifugation, Density Gradient
Chloramphenicol/pharmacology
Chromatography, Thin Layer
Coliphages/enzymology,growth & development
DNA, Viral
Escherichia coli/metabolism
Fatty Acids/metabolism
Glucose/metabolism
Lipid Metabolism
Lipopolysaccharides/metabolism
Micropore Filters
Muramidase/biosynthesis
Mutation
Phospholipids/metabolism
Polysaccharides, Bacterial/metabolism
Chemicals
Carbon Radioisotopes
DNA, Viral
Fatty Acids
Lipopolysaccharides
Phospholipids
Polysaccharides, Bacterial
Chloramphenicol
Muramidase
Glucose
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Loeb M R
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27 references, click to expand
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