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. 1979 Dec;32(3):917–924. doi: 10.1128/jvi.32.3.917-924.1979

Inhibition of initiation of bacteriophage T4 DNA replication by perturbation of Escherichia coli host membrane composition.

W M Huang
PMCID: PMC525940  PMID: 390167

Abstract

3-Decynoyl-N-acetylcysteamine (3-decynoyl-NAC) is an analog which specifically causes the immediate cessation of the biosynthesis of unsaturated fatty acids in Escherichia coli, whereas the synthesis of saturated fatty acids is actually stimulated. As a result, the cell membrane accumulates saturated fatty acids in its phospholipid. Addition of the inhibitor at the time of infection of E. coli by T4 phage had no effect on normal phage replication and development, implying that the synthesis of unsaturated fatty acids per se has little effect on T4 DNA replication. However, if the integrity and composition of the bacterial membrane was grossly perturbed by first treating the cells with the inhibitor for 60 min before infection, the proper initiation and the attainment of a rapid rate of T4 DNA synthesis were not observed. Under these conditions, a full complement of T4 early proteins was synthesized. The membrane associability of the known DNA delay proteins induced by wild-type T4 phage in the treated cells resembled that expected of a culture of untreated cells infected with a DNA delay mutant. When any one of three DNA delay mutants was used to infect 3-decynoyl-NAC-treated cells, T4 DNA replication was aborted. These findings suggest that some kind of specific interactions among the initiation proteins defined by the DNA delay mutants and the bacterial membrane may be necessary to facilitate the normal initiation and rapid rate of T4 DNA replication. A model for the involvement of the three different initiation proteins and the subsequent attainment of rapid DNA synthesis is discussed.

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Selected References

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

  1. Bolle A., Epstein R. H., Salser W., Geiduschek E. P. Transcription during bacteriophage T4 development: requirements for late messenger synthesis. J Mol Biol. 1968 Apr 28;33(2):339–362. doi: 10.1016/0022-2836(68)90193-9. [DOI] [PubMed] [Google Scholar]
  2. Brock D. J., Kass L. R., Bloch K. Beta-hydroxydecanoyl thioester dehydrase. II. Mode of action. J Biol Chem. 1967 Oct 10;242(19):4432–4440. [PubMed] [Google Scholar]
  3. Craine B. L., Rupert C. S. Identification of a biochemically unique DNA-membrane interaction involving the Escherichia coli origin of replication. J Bacteriol. 1978 Apr;134(1):193–199. doi: 10.1128/jb.134.1.193-199.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. EDGAR R. S., LIELAUSIS I. TEMPERATURE-SENSITIVE MUTANTS OF BACTERIOPHAGE T4D: THEIR ISOLATION AND GENETIC CHARACTERIZATION. Genetics. 1964 Apr;49:649–662. doi: 10.1093/genetics/49.4.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fralick J. A., Lark K. G. Evidence for the involvement of unsaturated fatty acids in initiating chromosome replication in Escherichia coli. J Mol Biol. 1973 Nov 5;80(3):459–475. doi: 10.1016/0022-2836(73)90416-6. [DOI] [PubMed] [Google Scholar]
  6. Gefter M. L. DNA replication. Annu Rev Biochem. 1975;44:45–78. doi: 10.1146/annurev.bi.44.070175.000401. [DOI] [PubMed] [Google Scholar]
  7. Huang W. M., Buchanan J. M. Synergistic interactions of T4 early proteins concerned with their binding to DNA. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2226–2230. doi: 10.1073/pnas.71.6.2226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Huang W. M. Membrane-associated proteins of T4-infected Escherichia coli. Virology. 1975 Aug;66(2):508–521. doi: 10.1016/0042-6822(75)90223-8. [DOI] [PubMed] [Google Scholar]
  9. Huang W. M. Positive regulation of T-even-phage DNA replication by the DNA-delay protein of gene 39. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 1):495–499. doi: 10.1101/sqb.1979.043.01.055. [DOI] [PubMed] [Google Scholar]
  10. Kass L. R., Bloch K. On the enzymatic synthesis of unsaturated fatty acids in Escherichia coli. Proc Natl Acad Sci U S A. 1967 Sep;58(3):1168–1173. doi: 10.1073/pnas.58.3.1168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kass L. R. The antibacterial activity of 3-decynoyl-n-acetylcysteamine. Inhibition in vivo of beta-hydroxydecanoyl thioester dehydrase. J Biol Chem. 1968 Jun 25;243(12):3223–3228. [PubMed] [Google Scholar]
  12. Kozinski A. W., Doermann A. H. Repetitive DNA replication of the incomplete genomes of phage T4 petite particles. Proc Natl Acad Sci U S A. 1975 May;72(5):1734–1738. doi: 10.1073/pnas.72.5.1734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McCarthy D., Minner C., Bernstein H., Bernstein C. DNA elongation rates and growing point distributions of wild-type phage T4 and a DNA-delay amber mutant. J Mol Biol. 1976 Oct 5;106(4):963–981. doi: 10.1016/0022-2836(76)90346-6. [DOI] [PubMed] [Google Scholar]
  14. Morris C. F., Sinha N. K., Alberts B. M. Reconstruction of bacteriophage T4 DNA replication apparatus from purified components: rolling circle replication following de novo chain initiation on a single-stranded circular DNA template. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4800–4804. doi: 10.1073/pnas.72.12.4800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nunn W. D., Cronan J. E., Jr Host membrane lipid synthesis is not required for successful phage T4 infection. Virology. 1976 Jan;69(1):332–335. doi: 10.1016/0042-6822(76)90221-x. [DOI] [PubMed] [Google Scholar]
  16. O'Farrell P. Z., Gold L. M., Huang W. M. The identification of prereplicative bacteriophage T4 proteins. J Biol Chem. 1973 Aug 10;248(15):5499–5501. [PubMed] [Google Scholar]
  17. Siegel P. J., Schaechter M. Bacteriophage T4 head maturation: release of progeny DNA from the host cell membrane. J Virol. 1973 Mar;11(3):359–367. doi: 10.1128/jvi.11.3.359-367.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Yegian C. D., Mueller M., Selzer G., Russo V., Stahl F. W. Properties of the DNA-delay mutants of bacteriophage T4. Virology. 1971 Dec;46(3):900–919. doi: 10.1016/0042-6822(71)90090-0. [DOI] [PubMed] [Google Scholar]

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