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. 1979 Oct;76(10):4877–4881. doi: 10.1073/pnas.76.10.4877

Isolation and identification of bacteriophage phi X174 prohead.

R Mukai, R K Hamatake, M Hayashi
PMCID: PMC413040  PMID: 159449

Abstract

The morphogenesis of bacteriophage phi X174 has been investigated by using an in vitro DNA synthesizing system. An extract of a B-mutant-infected cell is capable of synthesizing infectious phage in vitro when the B gene function is provided by the addition of an ammonium sulfate fraction of a C-mutant-infected-cell extract. This fraction contains the omega complex, a complex of phage-coded proteins with S = 108; the B-mutant extract does not. The purified omega complex, isolated from the C-mutant extract, caused the synthesis and encapsidation of viral DNA when added to B-mutant extract. The omega complex contains the B protein but it is the intact omega complex that functioned in the in vitro complementation of the B-mutant extract because other fractions containing B protein but no omega complex had little or no complementing activity. The results indicate that the omega complex is the phi X174 prohead. The B protein is not found in either the 132S or 114S phage particles but is an essential component of the prohead. This suggests that it may have a scaffolding function.

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Selected References

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

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  2. Eisenberg S., Griffith J., Kornberg A. phiX174 cistron A protein is a multifunctional enzyme in DNA replication. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3198–3202. doi: 10.1073/pnas.74.8.3198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fujisawa H., Hayashi M. Assembly of bacteriophage phi X174: identification of a virion capsid precursor and proposal of a model for the functions of bacteriophage gene products during morphogenesis. J Virol. 1977 Oct;24(1):303–313. doi: 10.1128/jvi.24.1.303-313.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fujisawa H., Hayashi M. Functions of gene C and gene D products of bacteriophage phi X 174. J Virol. 1977 Feb;21(2):506–515. doi: 10.1128/jvi.21.2.506-515.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fujisawa H., Hayashi M. Viral DNA-synthesizing intermediate complex isolated during assembly of bacteriophage phi X174. J Virol. 1976 Aug;19(2):409–415. doi: 10.1128/jvi.19.2.409-415.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gelfand D. H., Hayashi M. Electrophoretic characterization of phiX174-specific proteins. J Mol Biol. 1969 Sep 28;44(3):501–516. doi: 10.1016/0022-2836(69)90376-3. [DOI] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  9. Mukai R., Hayashi M. In vitro system that synthesizes circular viral DNA of bacteriophage phi X 174. J Virol. 1977 Jun;22(3):619–625. doi: 10.1128/jvi.22.3.619-625.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Mukai R., Hayashi M. Synthesis of infectious phiX-174 bacteriophage in vitro. Nature. 1977 Nov 24;270(5635):364–366. doi: 10.1038/270364a0. [DOI] [PubMed] [Google Scholar]
  11. Murialdo H., Becker A. Head morphogenesis of complex double-stranded deoxyribonucleic acid bacteriophages. Microbiol Rev. 1978 Sep;42(3):529–576. doi: 10.1128/mr.42.3.529-576.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Siden E. J., Hayashi M. Role of the gene beta-product in bacteriophage phi-X174 development. J Mol Biol. 1974 Oct 15;89(1):1–16. doi: 10.1016/0022-2836(74)90159-4. [DOI] [PubMed] [Google Scholar]
  13. Spindler K. R., Hayashi M. DNA synthesis in Escherichia coli cells infected with gene H mutants of bacteriophage phi X174. J Virol. 1979 Mar;29(3):973–982. doi: 10.1128/jvi.29.3.973-982.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Tonegawa S., Hayashi M. Intermediates in the assembly of phi X174. J Mol Biol. 1970 Mar 14;48(2):219–242. doi: 10.1016/0022-2836(70)90158-0. [DOI] [PubMed] [Google Scholar]

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