Summary
The RNA synthesis in non-viscous lysates containing the intact folded chromosome and cytoplasm fractions prepared from Escherichia coli has been examined in vitro. The RNA synthesis not only by chain extension but also by new chain initiation occurs in this system. While the RNA synthesis by chain extension takes place on the chromosome fraction alone (Pettijohn et al. 1970), an addition of the cytoplasm fraction is necessary for the synthesis by new chain initiations (de novo synthesis).
Analyses of the in vitro synthesized RNA by hybridization-competition and by sucrose gradient centrifugation show that 16S and 23S ribosomal RNAs account for about 40% of the total RNA products. The cytoplasm fraction is required for the de novo synthesis of ribosomal RNA at high relative rate. Guanosine tetraphosphate (ppGpp) does not specifically inhibit ribosomal RNA synthesis in this system.
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Atherly, A. G.: Ribonucleic acid regulation in permeabilized cells of E. coli capable of ribonucleic acid and protein synthesis. J. Bact. 118, 1186–1189, (1974)
Attardi, G., Huang, P. C., Kabat, S.: Recognition of ribosomal RNA sites in DNA, I. Analysis of the E. coli system. Proc. nat. Acad. Sci. (Wash.) 53, 1490–1497 (1965)
Cashel, M.: Inhibition of RNA polymerase by ppGpp, a nucleotide accumulated during the stringent response to amino acid starvation in Escherichia coli. Cold Spr. Harb. Symp. quant. Biol. 35, 407–413 (1970)
Dunn, J. J., Studier, F. W.: T7 early RNAs and Escherichia coli ribosomal RNAs are cut from large precursor RNAs in vivo by ribonuclease III. Proc. nat. Acad. Sci. (Wash.) 70, 3296–3300 (1973)
Edlin, G., Stent, G. S.: Nucleotide triphosphate pools and the regulation of RNA synthesis in E. coli. Proc. nat. Acad. Sci. (Wash.) 62, 475–482 (1969)
Gillespie, D., Spiegelman, S.: A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J. molec. Biol. 36, 1–14 (1965)
Haseltine W. A.: In vitro transcription, of Escherichia coli ribosomal RNA genes. Nature (Lond.) 235, 329–333 (1972)
Kennel, D.: Titration of the gene sites on DNA by DNA-RNA hybridization. II. The Escherichia coli chromosome. J. molec. Biol. 34, 85–103 (1968)
Lazzarini, R. A., Johnson, L. D.: Regulation of ribosomal RNA synthesis in cold-shocked E. coli. Nature (Lond.) New Biol. 243, 17–20 (1973)
Murooka, Y., Lazzarini, R. A.: In vitro synthesis of ribosomal ribonucleic acid by a deoxyribonucleic acid-protein complex isolated from Escherichia coli. J. biol. Chem. 248, 6248–6250 (1973)
Nikolaev N., Silengo, L., Schlessinger, D.: Synthesis of a large precursor to ribosomal RNA in a mutant of Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 70, 3361–3365 (1973)
Pace, N. P.: Structure and synthesis of the ribosomal ribonucleic acid of prokaryotes. Bact. Rev. 37, 562–603 (1973)
Pettijohn, D. E.: Ordered and preferential initiation of ribosomal RNA synthesis in vitro. Nature (Lond.) New Biol. 235, 204–206 (1972)
Pettijohn, D. E., Clarkson, K., Kossman, C. R., Stonington, O. G.: Synthesis of ribosomal RNA on a protein-DNA complex isolated from bacteria: a comparison of ribosomal RNA synthesis in vitro and in vivo. J. molec. Biol. 52, 281–300 (1970)
Pettijohn, D. E., Stonington, O. G., Kossman, C. R.: Chain termination of ribosomal RNA synthesis in vitro. Nature (Lond.) 228, 235–239 (1970)
Stonington, O. G., Pettijohn, D. E.: The folded genome of Escherichia coli isolated in a protein-DNA-RNA complex. Proc. nat. Acad. Sci. (Wash.) 68, 6–9 (1973)
Travers, A.: Control of ribosomal RNA synthesis in vitro. Nature (Lond.) 244, 15–17 (1973)
Travers, A., Kamen, R., Cashel, M.: The in vitro synthesis of ribosomal RNA. Cold Spr. Harb. Symp. quant. Biol. 35, 415–418 (1970)
Worcel, A., Burgi, E.: Properties of a membrane-attached form of the folded chromosome of Escherichia coli. J. molec. Biol. 82, 91–105 (1974)
Yankofsky, S. A., Spiegelman, S.: The identification of the ribosomal RNA cistron by sequence complementarity. I. Specificity of complex formation. Proc. nat. Acad. Sci. (Wash.) 48, 1069–1078 (1962)
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Communicated by H. G. Wittmann
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Muto, A. Preferential ribosomal RNA synthesis in the lysate of Escherichia coli . Molec. Gen. Genet. 138, 1–10 (1975). https://doi.org/10.1007/BF00268822
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DOI: https://doi.org/10.1007/BF00268822