Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1973 Mar;11(3):386–398. doi: 10.1128/jvi.11.3.386-398.1973

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

Robert L Swift 1, John S Wiberg 1
PMCID: PMC355114  PMID: 4570926

Abstract

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 μg/ml to 200 μg/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 − yi/yo), where yi and yo 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.

Full text

PDF
386

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BARRINGTON L. F., KOZLOFF L. M. Action of bacteriophage on isolated host cell walls. J Biol Chem. 1956 Dec;223(2):615–627. [PubMed] [Google Scholar]
  2. BOLLUM F. J. Thermal conversion of nonpriming deoxyribonucleic acid to primer. J Biol Chem. 1959 Oct;234:2733–2734. [PubMed] [Google Scholar]
  3. BONIFAS V., KELLENBERGER E. Etude de l'action des membranes du bactériophage T2 sur Escherichia coli. Biochim Biophys Acta. 1955 Mar;16(3):330–338. doi: 10.1016/0006-3002(55)90234-1. [DOI] [PubMed] [Google Scholar]
  4. BROWN D. D., KOZLOFF L. M. Morphological localization of the bacteriophage tail enzyme. J Biol Chem. 1957 Mar;225(1):1–11. [PubMed] [Google Scholar]
  5. Barbour S. D., Clark A. J. Biochemical and genetic studies of recombination proficiency in Escherichia coli. I. Enzymatic activity associated with recB+ and recC+ genes. Proc Natl Acad Sci U S A. 1970 Apr;65(4):955–961. doi: 10.1073/pnas.65.4.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Baumann L., Benz W. C., Wright A., Goldberg E. B. Inactivation of urea-treated phage T4 by phosphatidylglycerol. Virology. 1970 Jun;41(2):356–364. doi: 10.1016/0042-6822(70)90088-7. [DOI] [PubMed] [Google Scholar]
  7. Beppu T., Arima K. Properties of the colicin E 2 -induced degradation of deoxyribonucleic acid in Escherichia coli. J Biochem. 1971 Aug;70(2):263–271. doi: 10.1093/oxfordjournals.jbchem.a129638. [DOI] [PubMed] [Google Scholar]
  8. Changeux J. P., Thiéry J. On the mode of action of colicins: a model of regulation at the membrane level. J Theor Biol. 1967 Nov;17(2):315–318. doi: 10.1016/0022-5193(67)90175-0. [DOI] [PubMed] [Google Scholar]
  9. Chapman J. D., Swez J., Pollard E. C. Inhibition of gamma-ray-induced degradation of E. coli Bs-1 DNA by infection with T1, T2 and T4 bacteriophage. Nature. 1968 May 18;218(5142):690–693. doi: 10.1038/218690a0. [DOI] [PubMed] [Google Scholar]
  10. Corner T. R., Marquis R. E. Why do bacterial protoplasts burst in hypotonic solutions? Biochim Biophys Acta. 1969;183(3):544–558. doi: 10.1016/0005-2736(69)90168-0. [DOI] [PubMed] [Google Scholar]
  11. Dirksen M. L., Wiberg J. S., Koerner J. F., Buchanan J. M. EFFECT OF ULTRAVIOLET IRRADIATION OF BACTERIOPHAGE T2 ON ENZYME SYNTHESIS IN HOST CELLS. Proc Natl Acad Sci U S A. 1960 Nov;46(11):1425–1430. doi: 10.1073/pnas.46.11.1425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Duckworth D. H., Bessman M. J. Assay for the Killing Properties of T2 Bacteriophage and Their "Ghosts". J Bacteriol. 1965 Sep;90(3):724–728. doi: 10.1128/jb.90.3.724-728.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Duckworth D. H. Biological activity of bacteriophage ghosts and "take-over" of host functions by bacteriophage. Bacteriol Rev. 1970 Sep;34(3):344–363. doi: 10.1128/br.34.3.344-363.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Duckworth D. H. Inhibition of host deoxyribonucleic acid synthesis by T4 bacteriophage in the absence of protein synthesis. J Virol. 1971 Nov;8(5):754–758. doi: 10.1128/jvi.8.5.754-758.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Duckworth D. H. The metabolism of T4 phage ghost-infected cells. I. Macromolecular synthesis and ransport of nucleic acid and protein precursors. Virology. 1970 Mar;40(3):673–684. doi: 10.1016/0042-6822(70)90212-6. [DOI] [PubMed] [Google Scholar]
  16. Duckworth D. H., Winkler H. H. Metabolism of T4 bacteriophage ghost-infected cells. II. Do ghosts cause a generalized permeability change? J Virol. 1972 Jun;9(6):917–922. doi: 10.1128/jvi.9.6.917-922.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Edgar R. S., Lielausis I. Some steps in the assembly of bacteriophage T4. J Mol Biol. 1968 Mar 14;32(2):263–276. doi: 10.1016/0022-2836(68)90008-9. [DOI] [PubMed] [Google Scholar]
  18. Emrich J., Streisinger G. The role of phage lysozyme in the life cycle of phage T4. Virology. 1968 Nov;36(3):387–391. doi: 10.1016/0042-6822(68)90163-3. [DOI] [PubMed] [Google Scholar]
  19. FRASER D., JERREL E. A. The amino acid composition of T3 bacteriophage. J Biol Chem. 1953 Nov;205(1):291–295. [PubMed] [Google Scholar]
  20. FRENCH R. C., SIMINOVITCH L. The action of T2 bacteriophage ghosts on Escherichia coli B. Can J Microbiol. 1955 Dec;1(9):757–774. doi: 10.1139/m55-090. [DOI] [PubMed] [Google Scholar]
  21. Fabricant R., Kennell D. Inhibition of host protein synthesis during infection of Escherichi coli by bacteriophage T4. 3. Inhibition by ghosts. J Virol. 1970 Dec;6(6):772–781. doi: 10.1128/jvi.6.6.772-781.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Goldmark P. J., Linn S. An endonuclease activity from Escherichia coli absent from certain rec- strains. Proc Natl Acad Sci U S A. 1970 Sep;67(1):434–441. doi: 10.1073/pnas.67.1.434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. HERRIOTT R. M., BARLOW J. L. The protein coats or ghosts of coli phage T2. II. The biological functions. J Gen Physiol. 1957 Nov 20;41(2):307–331. doi: 10.1085/jgp.41.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Hercules K., Munro J. L., Mendelsohn S., Wiberg J. S. Mutants in a nonessential gene of bacteriophage T4 which are defective in the degradation of Escherichia coli deoxyribonucleic acid. J Virol. 1971 Jan;7(1):95–105. doi: 10.1128/jvi.7.1.95-105.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Holland E. M., Holland I. B. Induction of DNA breakdown and inhibition of cell division by colicin E2. Nature of some early steps in the process and properties of the E-2-specific nuclease system. J Gen Microbiol. 1970 Dec;64(2):223–239. doi: 10.1099/00221287-64-2-223. [DOI] [PubMed] [Google Scholar]
  26. KATZ W. VERGLEICHENDE UNTERSUCHUNGEN AN TEILCHENGEBUNDENEM UND FREIEM T2-LYSOZYM UND KRISTALLISIERUNG BEIDER ENZYME. Z Naturforsch B. 1964 Feb;19:129–133. [PubMed] [Google Scholar]
  27. King J. Assembly of the tail of bacteriophage T4. J Mol Biol. 1968 Mar 14;32(2):231–262. doi: 10.1016/0022-2836(68)90007-7. [DOI] [PubMed] [Google Scholar]
  28. Kutter E. M., Wiberg J. S. Biological effects of substituting cytosine for 5-hydroxymethylcytosine in the deoxyribonucleic acid of bacteriophage T4. J Virol. 1969 Oct;4(4):439–453. doi: 10.1128/jvi.4.4.439-453.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kutter E. M., Wiberg J. S. Degradation of cytosin-containing bacterial and bacteriophage DNA after infection of Escherichia coli B with bacteriophage T4D wild type and with mutants defective in genes 46, 47 and 56. J Mol Biol. 1968 Dec;38(3):395–411. doi: 10.1016/0022-2836(68)90394-x. [DOI] [PubMed] [Google Scholar]
  30. LEHMAN I. R., HERRIOTT R. M. The protein coats or ghosts or coliphage T2. III. Metabolic studies of Escherichia coli B infected with T2 bacteriophage ghosts. J Gen Physiol. 1958 May 20;41(5):1067–1082. doi: 10.1085/jgp.41.5.1067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. LEHMAN I. R., ROUSSOS G. G., PRATT E. A. The deoxyribonucleases of Escherichia coli. II. Purification and properties of a ribonucleic acid-inhibitable endonuclease. J Biol Chem. 1962 Mar;237:819–828. [PubMed] [Google Scholar]
  32. Lark K. G., Renger H. Initiation of DNA replication in Escherichia coli 15T-: chronological dissection of three physiological processes required for initiation. J Mol Biol. 1969 Jun 14;42(2):221–235. doi: 10.1016/0022-2836(69)90039-4. [DOI] [PubMed] [Google Scholar]
  33. Mitsui E., Mizuno D. Stabilization of colicin E2 by bovine serum albumin. J Bacteriol. 1969 Nov;100(2):1136–1137. doi: 10.1128/jb.100.2.1136-1137.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nomura M., Witten C., Mantei N., Echols H. Inhibition of host nucleic acid synthesis by bacteriophage T4: effect of chloramphenicol at various multiplicities of infection. J Mol Biol. 1966 May;17(1):273–278. doi: 10.1016/s0022-2836(66)80107-9. [DOI] [PubMed] [Google Scholar]
  35. PUCK T. T., LEE H. H. Mechanism of cell wall penetration by viruses. I. An increase in host cell permeability induced by bacteriophage infection. J Exp Med. 1954 May 1;99(5):481–494. doi: 10.1084/jem.99.5.481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pryme I. F., Joner P. E., Jensen H. B. The appearance of phage associated lysozyme in E. coli B cells immediately after infection with phage T2. Biochem Biophys Res Commun. 1969 Aug 15;36(4):676–681. doi: 10.1016/0006-291x(69)90359-3. [DOI] [PubMed] [Google Scholar]
  37. Ringrose P. Sedimentation analysis of DNA degradation products resulting from the action of colicin E2 on Escherichia coli. Biochim Biophys Acta. 1970 Aug 8;213(2):320–334. doi: 10.1016/0005-2787(70)90040-7. [DOI] [PubMed] [Google Scholar]
  38. STENT G. S., WOLLMAN E. L. On the two step nature of bacteriophage absorption. Biochim Biophys Acta. 1952 Mar;8(3):260–269. doi: 10.1016/0006-3002(52)90041-3. [DOI] [PubMed] [Google Scholar]
  39. Silver S., Levine E., Spielman P. M. Cation fluxes and permeability changes accompanying bacteriophage infection of Escherichia coli. J Virol. 1968 Aug;2(8):763–771. doi: 10.1128/jvi.2.8.763-771.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Simon L. D., Anderson T. F. The infection of Escherichia coli by T2 and T4 bacteriophages as seen in the electron microscope. I. Attachment and penetration. Virology. 1967 Jun;32(2):279–297. doi: 10.1016/0042-6822(67)90277-2. [DOI] [PubMed] [Google Scholar]
  41. Simon L. D., Swan J. G., Flatgaard J. E. Functional defects in T4 bacteriophages lacking the gene 11 and gene 12 products. Virology. 1970 May;41(1):77–90. doi: 10.1016/0042-6822(70)90056-5. [DOI] [PubMed] [Google Scholar]
  42. Swift R. L., Wiberg J. S. Bacteriophage T4 inhibits colicin E2-induced degradation of Escherichia coli deoxyribonucleic acid. I. Protein synthesis-dependent inhibition. J Virol. 1971 Sep;8(3):303–310. doi: 10.1128/jvi.8.3.303-310.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. WIBERG J. S., DIRKSEN M. L., EPSTEIN R. H., LURIA S. E., BUCHANAN J. M. Early enzyme synthesis and its control in E. coli infected with some amber mutants of bacteriophage T4. Proc Natl Acad Sci U S A. 1962 Feb;48:293–302. doi: 10.1073/pnas.48.2.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ward C. B., Glaser D. A. Analysis of the chloramphenicol-sensitive and chloramphenicol-resistant steps in the initiation of DNA synthesis in E. coli B-r. Proc Natl Acad Sci U S A. 1969 Nov;64(3):905–912. doi: 10.1073/pnas.64.3.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Warner H. R., Snustad P., Jorgensen S. E., Koerner J. F. Isolation of bacteriophage T4 mutants defective in the ability to degrade host deoxyribonucleic acid. J Virol. 1970 Jun;5(6):700–708. doi: 10.1128/jvi.5.6.700-708.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Warren R. J., Bose S. K. Bacteriophage-induced inhibition of host functions. I. Degradation of Escherichia coli deoxyribonucleic acid after T4 infection. J Virol. 1968 Apr;2(4):327–334. doi: 10.1128/jvi.2.4.327-334.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wiberg J. S. Amber mutants of bacteriophage T4 defective in deoxycytidine diphosphatase and deoxycytidine triphosphatase. On the role of 5-hydroxymethylcytosine in bacteriophage deoxyribonucleic acid. J Biol Chem. 1967 Dec 25;242(24):5824–5829. [PubMed] [Google Scholar]
  48. Wiberg J. S. Mutants of bacteriophage T4 unable to cause breakdown of host DNA. Proc Natl Acad Sci U S A. 1966 Mar;55(3):614–621. doi: 10.1073/pnas.55.3.614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wilson J. H., Luftig R. B., Wood W. B. Interaction of bacteriophage T4 tail fiber components with a lipopolysaccharide fraction from Escherichia coli. J Mol Biol. 1970 Jul 28;51(2):423–434. doi: 10.1016/0022-2836(70)90152-x. [DOI] [PubMed] [Google Scholar]
  50. Winkler H. H., Duckworth D. H. Metabolism of T4 bacteriophage ghost-infected cells: effect of bacteriophage and ghosts on the uptake of carbohydrates in Escherichia coli B. J Bacteriol. 1971 Jul;107(1):259–267. doi: 10.1128/jb.107.1.259-267.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES