Summary
A cell free protein synthesizing system, derived from E. coli, is shown to be a quantitative assay system for messenger RNA extracted from B. subtilis infected with bacteriophage SPO1. DNA-directed protein synthesis in this system is shown to be limited mostly to those proteins whose messages are contained in early RNA. A phage induced enzyme, dCMP deaminase, is shown to be dependent on appearance of a class of mRNA made in vivo in response to new initiations of transcription dependent on prior synthesis of phage induced protein. Control of the enzyme synthesized in the cell free system is contrasted with in vivo control, and an estimate of “read-through” by RNA polymerase in vitro is presented.
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Abbreviations
- SDS:
-
sodium dodecyl sulfate
- CM:
-
chloramphenicol
References
Fairbanks, G., Levinthal, C., Reeder, R.H.: Analysis of 14C-labeled proteins by disc electrophoresis. Biochem. biophys. Res. Commun. 20, 393–399 (1965)
Gage, L.P., Geiduschek, E.P.: RNA synthesis during bacteriophage SPO1 development: six classes of SPO1 RNA. J. molec. Biol. 57, 279–300 (1971a)
Gage, L.P., Geiduschek, E.P.: RNA synthesis during bacteriophage SPO1 development. II: Some modulations and prerequisites of the transcription program. Virology 44, 200–210 (1971b)
Geiduschek, E.P., Brody, E.N., Wilson, D.L.: Some aspects of RNA transcription. In: Molecular associations in biology, ed. Pullman, B., p. 163–182. New York: Academic Press 1968
Gold, L.M., Schweiger, M.: control of β-glucosyltransferase and lysozyme synthesis during T4 deoxyribonucleic acid-dependent ribonucleic acid and protein synthesis in vitro. J. biol. Chem. 245, 2255–2258 (1970)
Gold, L.M., Schweiger, M.: In: Methods in enzymology vol. 20, eds. Moldave, K. and Grossman, L., p. 537–452. New York: Academic Press 1971
Leammli, U.K.: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (Lond.) 207, 680–685 (1970)
Levinthal, C., Hosoda, J., Shub, D.A.: Control of protein synthesis after phage infection. In: Molecular biology of viruses, eds. Colter, J.S. and Paranchych, W., p. 71–87. New York: Academic Press 1967
O'Farrell, P.Z., Gold, L.M.: Transcription and translation of pre-replicative bacteriophage T4 genes in vitro. J. biol. Chem. 248, 5502–5511 (1973)
Okubo, S., Strauss, B.S., Stodolsky, M.: The possible role of recombination in the infection of competent Bacillus subtilis by bacteriophage nucleic acid. Virology 24, 552–562 (1964)
Schweiger, M., Gold, L.M.: Escherichia coli and Bacillus subtilis phage deoxyribonucleic acid-directed deoxycytidylate deaminase synthesis in Escherichia coli extracts. J. biol. Chem. 245, 5022–5025 (1970)
Shub, D.A.: Stability of messenger RNA during bacteriophage development. Ph.D. Thesis, Massachusetts Institute of Technology (1966)
Shub, D.A., Johnson, G.G.: Bacteriophage SPO1 DNA- and RNA-directed protein synthesis in vitro: the effect of TFl, a template-selective transcription inhibitor. Molec. gen. Genet. 137, 161–169 (1975)
Thomas, C.A., Abelson, J.: The isolation and characterization of DNA from bacteriophage. In: Procedures in nucleic acid research, eds. Cantoni, G.L. Davies, D.R., p. 553–561. New York: Harper & Row 1966
Truffaut, N., Revet, B., Soulie, M.: Étude Comparative des DNA de Phages 2C, SP8*, SP82, Φe, SPO1 et SP50. Europ. J. Biochem. 15, 391–400 (1970)
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Communicated by W. Maas
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Shub, D.A. Bacteriophage SPO1 DNA- and RNA-directed protein synthesis in vitro: Comparison with in vivo control. Molec. gen. Genet. 137, 171–180 (1975). https://doi.org/10.1007/BF00341683
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DOI: https://doi.org/10.1007/BF00341683