Bacteriophage T4 inhibits colicin E2-induced degradation of Escherichia coli deoxyribonucleic acid. II. Inhibition by T4 ghosts and by T4 in the absence of protein synthesis.
The deoxyribonucleic acid (DNA) of Escherichia coli B is converted by colicin E2 to products soluble in cold trichloroacetic acid; we showed previously that this DNA degradation (hereafter termed solubilization) is subject to inhibition by infection with phage T4 and that at least two modes of inhibition can be differentiated on the basis of their sensitivity to chloramphenicol (CM). This report deals exclusively with the inhibition of E2 produced by T4, or T4 ghosts, in the absence of protein synthesis. The following observations are described. (i) The stage of T4 infection that inhibits E2 occurs after reversible adsorption of the phage to the bacterial surface, but probably prior to injection of T4 DNA into the cell's interior. (ii) The extent of inhibition increases as the T4 multiplicity is increased; however, the fraction of bacterial DNA that eventually is solubilized is virtually independent of the phage multiplicity. (iii) Phage ghosts (DNA-less phage particles) possess an approximately 15-fold greater inhibitory capacity toward E2 than do intact phage; however, because highly purified T4 (completely freed of ghost contamination) still inhibit E2, we discount the possibility that preparations of "intact phage" inhibit exclusively by virtue of contaminating ghosts. (iv) T4 infection does not liberate an extracellular inactivator of E2. In fact, infection with sufficiently high multiplicities of T4 produces a supernatant factor that protects E2 from nonspecific inactivation at 37 C. This protective factor does not interfere with the colicin's ability to induce DNA solubilization. (v) Inhibition of E2 occurs even when phage are added well after initiation of DNA solubilization by E2, suggesting that a late stage of E2 action is the target of inhibition by T4 infection. (vi) Increasing the CM concentration from 50 mug/ml to 200 mug/ml appears to reduce the inhibition appreciably; however, this can be attributed to an enhancement by CM of the rate of E2-induced DNA solubilization. (vii) The same degree of inhibition of E2 by T4 seen in CM is observed when CM is replaced by puromycin or rifampin. (viii) Others have shown that raising the multiplicity of E2 increases the rate of DNA solubilization. We find that the fractional inhibition (i), [i = (1 - y(i)/y(o)), where y(i) and y(o) represent the inhibited and uninhibited rates of solubilization of DNA, respectively], produced by a given T4 multiplicity is independent of the multiplicity of E2 and hence is independent of the rate of DNA solubilization induced by E2.
D000327:Adsorption; D001426:Bacterial Proteins / Q000096:biosynthesis*; D002247:Carbon Isotopes; D002701:Chloramphenicol / Q000494:pharmacology; D003087:Colicins / Q000494:pharmacology*; D003090:Coliphages / Q000254:growth & development; D004267:DNA Viruses / Q000254:growth & development; D004269:DNA, Bacterial / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D007930:Leucine; D009154:Mutation; D009691:Nucleic Acid Denaturation; D011691:Puromycin / Q000494:pharmacology; D012293:Rifampin / Q000494:pharmacology; D012995:Solubility; D014316:Tritium; D014498:Uracil; D014764:Viral Proteins / Q000494:pharmacology*; D014779:Virus Replication
D001426:Bacterial Proteins; D002247:Carbon Isotopes; D003087:Colicins; D004269:DNA, Bacterial; D014764:Viral Proteins; D014316:Tritium; D011691:Puromycin; D014498:Uracil; D002701:Chloramphenicol; D007930:Leucine; D012293:Rifampin