Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1982 Jun;42(3):1099–1107. doi: 10.1128/jvi.42.3.1099-1107.1982

Minor coat protein composition and location of the A protein in bacteriophage f1 spheroids and I-forms.

J Lopez, R E Webster
PMCID: PMC256948  PMID: 7097858

Abstract

The filamentous bacteriophage f1 can be transformed into a spherical particle (spheroid) or an intermediate shortened filament with a flared end (I-forms) by exposure to a chloroform-water interface at 22 or 4 degrees C, respectively. The protein composition of bacteriophage f1 spheroids and I-forms was examined by separating the proteins from the purified. [35S]cysteine-labeled particles by sodium dodecyl sulfate-urea-polyacrylamide gel electrophoresis. Quantitation of the radioactivity on the gels showed that I-forms and spheroids contain the same complement of minor coat proteins as do untreated f1 phage. This composition is unchanged after removal of the DNA, either by digestion with micrococcal nuclease or by centrifugation of the particles through CsCl density gradients, indicating that none of the minor coat proteins is held in the particles solely through an interaction with the DNA. We also examined the location of the A protein in I-forms by decoration with ferritin-conjugated antibodies and examination under the electron microscope and found that the A protein is located specifically at the flared end of the I-form particle, through which the DNA is extruded and at which contraction into spheroids begins. The implications of these results with regard to the orientation of the DNA within the capsid and the process of infection are discussed.

Full text

PDF
1099

Images in this article

1102Image
on p.1102
1103Image
on p.1103
1106Image
on p.1106

Selected References

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

  1. Burke J. M., Novotny C. P., Fives-Taylor P. Defective F pili and other characteristics of Flac and Hfr Escherichia coli mutants resistant to bacteriophage R17. J Bacteriol. 1979 Nov;140(2):525–531. doi: 10.1128/jb.140.2.525-531.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Caro L. G., Schnös M. The attachment of the male-specific bacteriophage F1 to sensitive strains of Escherichia coli. Proc Natl Acad Sci U S A. 1966 Jul;56(1):126–132. doi: 10.1073/pnas.56.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Date T., Inuzuka M., Tomoeda M. Purification and characterization of F pili from Escherichia coli. Biochemistry. 1977 Dec 13;16(25):5579–5585. doi: 10.1021/bi00644a030. [DOI] [PubMed] [Google Scholar]
  4. Frank H., Day L. A. Electron microscopic observations on fd bacteriophage, its alkali denaturation products and its DNA. Virology. 1970 Sep;42(1):144–154. doi: 10.1016/0042-6822(70)90247-3. [DOI] [PubMed] [Google Scholar]
  5. Goldsmith M. E., Konigsberg W. H. Adsorption protein of the bacteriophage fd: isolation, molecular properties, and location in the virus. Biochemistry. 1977 Jun 14;16(12):2686–2694. doi: 10.1021/bi00631a016. [DOI] [PubMed] [Google Scholar]
  6. Grant R. A., Lin T. C., Konigsberg W., Webster R. E. Structure of the filamentous bacteriophage fl. Location of the A, C, and D minor coat proteins. J Biol Chem. 1981 Jan 10;256(1):539–546. [PubMed] [Google Scholar]
  7. Gray C. W., Brown R. S., Marvin D. A. Adsorption complex of filamentous fd virus. J Mol Biol. 1981 Mar 15;146(4):621–627. doi: 10.1016/0022-2836(81)90050-4. [DOI] [PubMed] [Google Scholar]
  8. Griffith J., Manning M., Dunn K. Filamentous bacteriophage contract into hollow spherical particles upon exposure to a chloroform-water interface. Cell. 1981 Mar;23(3):747–753. doi: 10.1016/0092-8674(81)90438-4. [DOI] [PubMed] [Google Scholar]
  9. Jacobson A. Role of F pili in the penetration of bacteriophage fl. J Virol. 1972 Oct;10(4):835–843. doi: 10.1128/jvi.10.4.835-843.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lin T. C., Webster R. E., Konigsberg W. Isolation and characterization of the C and D proteins coded by gene IX and gene VI in the filamentous bacteriophage fl and fd. J Biol Chem. 1980 Nov 10;255(21):10331–10337. [PubMed] [Google Scholar]
  11. Lyons L. B., Zinder N. D. The genetic map of the filamentous bacteriophage f1. Virology. 1972 Jul;49(1):45–60. doi: 10.1016/s0042-6822(72)80006-0. [DOI] [PubMed] [Google Scholar]
  12. Makino S., Woolford J. L., Jr, Tanford C., Webster R. E. Interaction of deoxycholate and of detergents with the coat protein of bacteriophage f1. J Biol Chem. 1975 Jun 10;250(11):4327–4332. [PubMed] [Google Scholar]
  13. Manning M., Chrysogelos S., Griffith J. Mechanism of coliphage M13 contraction: intermediate structures trapped at low temperatures. J Virol. 1981 Dec;40(3):912–919. doi: 10.1128/jvi.40.3.912-919.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nelson F. K., Friedman S. M., Smith G. P. Filamentous phage DNA cloning vectors: a noninfective mutant with a nonpolar deletion in gene III. Virology. 1981 Jan 30;108(2):338–350. doi: 10.1016/0042-6822(81)90442-6. [DOI] [PubMed] [Google Scholar]
  15. Ohkawa I., Webster R. E. The orientation of the major coat protein of bacteriophage f1 in the cytoplasmic membrane of Escherichia coli. J Biol Chem. 1981 Oct 10;256(19):9951–9958. [PubMed] [Google Scholar]
  16. Pratt D., Tzagoloff H., Beaudoin J. Conditional lethal mutants of the small filamentous coliphage M13. II. Two genes for coat proteins. Virology. 1969 Sep;39(1):42–53. doi: 10.1016/0042-6822(69)90346-8. [DOI] [PubMed] [Google Scholar]
  17. Rossomando E. F. Studies on the structural polarity of bacteriophage f1. Virology. 1970 Nov;42(3):681–687. doi: 10.1016/0042-6822(70)90313-2. [DOI] [PubMed] [Google Scholar]
  18. Rossomando E. F., Zinder N. D. Studies on the bacteriophage fl. I. Alkali-induced disassembly of the phage into DNA and protein. J Mol Biol. 1968 Sep 28;36(3):387–399. doi: 10.1016/0022-2836(68)90163-0. [DOI] [PubMed] [Google Scholar]
  19. Shen C. K., Ikoku A., Hearst J. E. A specific DNA orientation in the filamentous bacteriophage fd as probed by psoralen crosslinking and electron microscopy. J Mol Biol. 1979 Jan 15;127(2):163–175. doi: 10.1016/0022-2836(79)90237-7. [DOI] [PubMed] [Google Scholar]
  20. Simons G. F., Konings R. N., Schoenmakers J. G. Genes VI, VII, and IX of phage M13 code for minor capsid proteins of the virion. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4194–4198. doi: 10.1073/pnas.78.7.4194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Smilowitz H. Bacteriophage f1 infection: fate of the parental major coat protein. J Virol. 1974 Jan;13(1):94–99. doi: 10.1128/jvi.13.1.94-99.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Webster R. E., Grant R. A., Hamilton L. A. Orientation of the DNA in the filamentous bacteriophage f1. J Mol Biol. 1981 Oct 25;152(2):357–374. doi: 10.1016/0022-2836(81)90247-3. [DOI] [PubMed] [Google Scholar]
  23. Wickner W. Asymmetric orientation of a phage coat protein in cytoplasmic membrane of Escherichia coli. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4749–4753. doi: 10.1073/pnas.72.12.4749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Woolford J. L., Jr, Steinman H. M., Webster R. E. Adsorption protein of bacteriophage fl: solubilization in deoxycholate and localization in the fl virion. Biochemistry. 1977 Jun 14;16(12):2694–2700. doi: 10.1021/bi00631a017. [DOI] [PubMed] [Google Scholar]

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

RESOURCES