Abstract
An in vitro system was developed for the study of the initial stages of bacteriophage phi chi 174 infection. Escherichia coli C cells were incubated with 20% sucrose and then subjected to cold osmotic shock in 5 mM MgSO4. The concentrated supernatant shock fluid inactivated phi chi 174 with the same kinetics and requirements as for normal infection. Shock fluids prepared from phi chi 174-resistant strains of E. coli did not show this effect. The 114S phage were initially converted into 70S particles, the process termed "eclipse". These structurally altered phages then attached to a component of the shock fluid, producing fast-sedimenting complexes, and eventually released at least a part of their DNA into the medium. The fast-sedimenting complex could be radioactively labeled with oleic acid. Radioactivity was found to co-chromatograph with both biological activity and the majority of the high-molecular-weight carbohydrates present in the shock fluid. It is concluded that E. coli C osmotic shock fluid contains isolated phi chi 174-specific receptor sites composed of lipopolysaccharides. This system conveniently separates the early stages of phage phi chi 174 infection from the intracellular events.
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Selected References
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- Beswick F. M., Lunt M. R. Adsorption of bacteriophage phiX174 to isolated bacterial cell walls. J Gen Virol. 1972 Sep;16(3):381–390. doi: 10.1099/0022-1317-16-3-381. [DOI] [PubMed] [Google Scholar]
- Brown D. T., MacKenzie J. M., Bayer M. E. Mode of host cell penetration by bacteriophage phi X174. J Virol. 1971 Jun;7(6):836–846. doi: 10.1128/jvi.7.6.836-846.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GUTHRIE G. D., SINSHEIMER R. L. Observations on the infection of bacterial protoplasts with the deoxyribonucleic acid of bacteriophage phi X174. Biochim Biophys Acta. 1963 Jun 25;72:290–297. [PubMed] [Google Scholar]
- Hutchison C. A., 3rd, Sinsheimer R. L. The process of infection with bacteriophage phi-X174. X. Mutations in a phi-X Lysis gene. J Mol Biol. 1966 Jul;18(3):429–447. doi: 10.1016/s0022-2836(66)80035-9. [DOI] [PubMed] [Google Scholar]
- Incardona N. L., Blonski R., Feeney W. Mechanism of adsorption and eclipse of bacteriophage phi X174. I. In vitro conformational change under conditions of eclipse. J Virol. 1972 Jan;9(1):96–101. doi: 10.1128/jvi.9.1.96-101.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Incardona N. L., Selvidge L. Mechanism of adsorption and eclipse of bacteriophage phi X174. II. Attachment and eclipse with isolated Escherichia coli cell wall lipopolysaccharide. J Virol. 1973 May;11(5):775–782. doi: 10.1128/jvi.11.5.775-782.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knippers R., Salivar W. O., Newbold J. E., Sinsheimer R. L. The process of infection with bacteriophage phiX174. XXVI. Transfer of the parental DNA of bacteriophage phiX174 into progeny bacteriophage particles. J Mol Biol. 1969 Feb 14;39(3):641–654. doi: 10.1016/0022-2836(69)90150-8. [DOI] [PubMed] [Google Scholar]
- Knippers R., Sinsheimer R. L. Process of infection with bacteriophage phiX174. XX. Attachment of the parental DNA of bacteriophage phiX174 to a fast-sedimenting cell component. J Mol Biol. 1968 May 28;34(1):17–29. doi: 10.1016/0022-2836(68)90231-3. [DOI] [PubMed] [Google Scholar]
- 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]
- Lindberg A. A. Bacteriophage receptors. Annu Rev Microbiol. 1973;27:205–241. doi: 10.1146/annurev.mi.27.100173.001225. [DOI] [PubMed] [Google Scholar]
- Neu H. C., Heppel L. A. The release of enzymes from Escherichia coli by osmotic shock and during the formation of spheroplasts. J Biol Chem. 1965 Sep;240(9):3685–3692. [PubMed] [Google Scholar]
- Newbold J. E., Sinsheimer R. L. Process of infection with bacteriophage phi-X174. XXXIV. Kinetic of the attachment and eclipse steps of the infection. J Virol. 1970 Apr;5(4):427–431. doi: 10.1128/jvi.5.4.427-431.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Newbold J. E., Sinsheimer R. L. The process of infection with bacteriophage phiX174. XXXII. Early steps in the infection process: attachment, eclipse and DNA penetration. J Mol Biol. 1970 Apr 14;49(1):49–66. doi: 10.1016/0022-2836(70)90375-x. [DOI] [PubMed] [Google Scholar]
- Nossal N. G., Heppel L. A. The release of enzymes by osmotic shock from Escherichia coli in exponential phase. J Biol Chem. 1966 Jul 10;241(13):3055–3062. [PubMed] [Google Scholar]
- Salivar W. O., Sinsheimer R. L. Intracellular location and number of replicating parental DNA molecules of bacteriophages lambda and phi-X174. J Mol Biol. 1969 Apr 14;41(1):39–65. doi: 10.1016/0022-2836(69)90124-7. [DOI] [PubMed] [Google Scholar]
- Yarus M. J., Sinsheimer R. L. The process of infection with bacteriophage phiX174. 8. Evidence for an essential bacterial "site". J Virol. 1967 Feb;1(1):135–144. doi: 10.1128/jvi.1.1.135-144.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
