Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1975 Sep;16(3):685–695. doi: 10.1128/jvi.16.3.685-695.1975

Proteins of bacteriophage phi6.

J F Sinclair, A Tzagoloff, D Levine, L Mindich
PMCID: PMC354716  PMID: 1159897

Abstract

We investigated the protein composition of the lipid-containing bacteriophage phi 6. We also studied the synthesis of phage-specific proteins in the host bacterium Pseudomonas phaseolicola HB10Y. The virion was found to contain 10 proteins of the following molecular weights: P1, 93,000; P2, 88,000; P3, 84,000; P4, 36,800; P5, 24,000; P6, 21,000; P7, 19,900; P8, 10,500; P9, 8,700; and P10, less than 6,000. Proteins P3, P9, and P10 were completely extracted from the virion with 1% Triton X-100. Protein P6 was partially extracted. Proteins P8 and P9 were purified by column chromatography. The amino acid composition of P9 was determined and was found to lack methionine. Labeling of viral proteins with [35S]methionine in infected cells indicated that proteins P5, P9, P10, and P11 lacked methionine. Treatment of host cells with UV light before infection allowed the synthesis of P1, P2, P4, and P7; however, the extent of viral protein synthesis fell off exponentially with increasing delay time between irradiation and infection. Treatment of host cells with rifampin during infection allowed preferential synthesis of viral proteins, but the extent of synthesis also fell off exponentially with increasing delay time between the addition of rifampin and the addition of radioactive amino acids. All of the virion proteins were seen in gels prepared from rifampin-treated infected cells. In addition, two proteins, P11 and P12, were observed; their molecular weights were 25,200 and 20,100, respectively. Proteins P1, P2, P4, and P7 were synthesized early, whereas the rest began to increase at 45 min post-infection.

Full text

PDF
685

Images in this article

Selected References

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

  1. Camerini-Otero R. D., Franklin R. M. Structure and synthesis of a lipid-containing bacteriophage. XII. The fatty acids and lipid content of bacteriophage PM2. Virology. 1972 Aug;49(2):385–393. doi: 10.1016/0042-6822(72)90491-6. [DOI] [PubMed] [Google Scholar]
  2. Chamberlin M., McGrath J., Waskell L. New RNA polymerase from Escherichia coli infected with bacteriophage T7. Nature. 1970 Oct 17;228(5268):227–231. doi: 10.1038/228227a0. [DOI] [PubMed] [Google Scholar]
  3. Ellis L. F., Schlegel R. A. Electron microscopy of Pseudomonas phi 6 bacteriophage. J Virol. 1974 Dec;14(6):1547–1551. doi: 10.1128/jvi.14.6.1547-1551.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Espejo R. T., Canelo E. S. Properties of bacteriophage PM2: a lipid-containing bacterial virus. Virology. 1968 Apr;34(4):738–747. doi: 10.1016/0042-6822(68)90094-9. [DOI] [PubMed] [Google Scholar]
  5. Espejo R. T., Canelo E. S., Sinsheimer R. L. DNA of bacteriophage PM2: a closed circular double-stranded molecule. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1164–1168. doi: 10.1073/pnas.63.4.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grinsted J., Saunders J. R., Ingram L. C., Sykes R. B., Richmond M. H. Properties of a R factor which originated in Pseudomonas aeruginosa 1822. J Bacteriol. 1972 May;110(2):529–537. doi: 10.1128/jb.110.2.529-537.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hinnen R., Schäfer R., Franklin R. M. Structure and synthesis of a lipid-containing bacteriophage. Preparation of virus and localization of the structural proteins. Eur J Biochem. 1974 Dec 16;50(1):1–14. doi: 10.1111/j.1432-1033.1974.tb03867.x. [DOI] [PubMed] [Google Scholar]
  8. Hosoda J., Levinthal C. Protein synthesis by Escherichia coli infected with bacteriophage T4D. Virology. 1968 Apr;34(4):709–727. doi: 10.1016/0042-6822(68)90092-5. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. O'Farrell P. Z., Gold L. M. Bacteriophage T4 gene expression. Evidence for two classes of prereplicative cistrons. J Biol Chem. 1973 Aug 10;248(15):5502–5511. [PubMed] [Google Scholar]
  11. Semancik J. S., Vidaver A. K., Van Etten J. L. Characterization of segmented double-helical RNA from bacteriophage phi6. J Mol Biol. 1973 Aug 25;78(4):617–625. doi: 10.1016/0022-2836(73)90283-0. [DOI] [PubMed] [Google Scholar]
  12. Shatkin A. J. Viruses with segmented ribonucleic acid genomes: multiplication of influenza versus reovirus. Bacteriol Rev. 1971 Sep;35(3):250–266. doi: 10.1128/br.35.3.250-266.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  14. Studier F. W., Maizel J. V., Jr T7-directed protein synthesis. Virology. 1969 Nov;39(3):575–586. doi: 10.1016/0042-6822(69)90105-6. [DOI] [PubMed] [Google Scholar]
  15. Van Etten J. L., Vidaver A. K., Koski R. K., Semancik J. S. RNA polymerase activity associated with bacteriophage phi 6. J Virol. 1973 Sep;12(3):464–471. doi: 10.1128/jvi.12.3.464-471.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Vidaver A. K., Koski R. K., Van Etten J. L. Bacteriophage phi6: a Lipid-Containing Virus of Pseudomonas phaseolicola. J Virol. 1973 May;11(5):799–805. doi: 10.1128/jvi.11.5.799-805.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Watanabe Y., Millward S., Graham A. F. Regulation of transcription of the Reovirus genome. J Mol Biol. 1968 Aug 28;36(1):107–123. doi: 10.1016/0022-2836(68)90223-4. [DOI] [PubMed] [Google Scholar]
  18. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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

RESOURCES