Summary
-
1.
A native DNA fraction was isolated from the young vegetative mycelium of Streptomyces griseus strain No. 52-1 and was compared with a similar fraction of Escherichia coli B. The endogenous RNA polymerase activities of these DNA fractions were examined.
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2.
Determining the base composition of the 14C labelled RNA synthesized in vitro, it was found that the conversion of 14C UTP into CMP residue of RNA is negligible in the DNA fraction of S. griseus, while it is significant in the DNA fraction of E. coli. By the proof of density gradient centrifugation, the RNA synthesized in vitro was mainly of messenger size in both species.
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3.
Template activity of the native DNA fractions was investigated in the presence of isolated E. coli RNA polymerase. When measuring the endogenous RNA polymerase and template activities of the DNA complexes at lower and higher ionic strength it was established that DNA complexes of E. coli under both ionic conditions behaved essentially as free DNA strands did, while endogenous and exogenous RNA polymerase activities in DNA complexes of S. griseus are inhibited in medium of lower ionic strength. Our results indicate the presence of some additional compound in the S. griseus aggregate which is absent in E. coli. This compound could function at lower ionic strength while it would be inactivated at higher ionic strength.
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Abbreviations
- TMA:
-
medium: 0.01 M tris-HCl pH 7.8
- 0.01 M:
-
magnesium acetate
- 0.06 M:
-
KCl
- 0.006 M:
-
mercaptoethanol
- PCA:
-
perchloric acid
References
Butler, J. A. V., Godson, G. N.: Biosynthesis of nucleic acids in Bacillus megaterium. 1. The isolation of a nuclear material. Biochem. J. 88, 176–182 (1963).
Bu'Lock, J. D.: The biosynthesis of natural products. An introduction to secondary metabolism. London: McGraw-Hill 1965.
Chamberlin, M., Berg, P.: Deoxyribonucleic acid-directed synthesis of ribonucleic acid by an enzyme from Escherichia coli. Proc. nat. Acad. Sci. (Wash.) A 48, 81–94 (1961).
Fuchs, E., Millette, R. L., Zillig, W., Walter, G.: Influence of salts on RNA synthesis by DNA-dependent RNA polymerase from Escherichia coli. Europ. J. Biochem. 3, 183–193 (1967).
Godson, G. N., Butler, J. A. V.: Biosynthesis of nucleic acids in Bacillus megaterium. 4. Roles of the “nuclear”, cytoplasmic and cytoplasmic membrane components of the cell in the biosynthesis of ribonucleic acid. Biochem. J. 93, 573–583 (1964).
Hopwood, D. A., Glauert, A. M.: Observations on the chromatinic bodies of Streptomyces coelicolor. J. biophys. biochem. Cytol. 8, 257–265 (1960).
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the Folin phenol reagent. J. biol. Chem. 193, 265–275 (1951).
Lubin, M.: Observation on the structure of RNA polymerase and its attachment to DNA. J. molec. Biol. 39, 219–233 (1969).
Matthaei, J. H., Nirenberg, M. W.: Characteristics and stabilization of DNAase sensitive protein synthes is in E. coli extracts. Proc. nat. Acad. Sci. (Wash.) A 47, 1580–1588 (1961).
McCarthy, B. J., Britten, R. J.: The synthesis of ribosomes in E. coli. 1. The incorporation of 14C uracil into the metabolic pool and RNA. Biophys. J. 2, 35–47 (1962).
Mizuno, S., Whiteley, H. R.: Nuclear fraction of Bacillus subtilis as a template for ribonucleic acid synthesis. J. Bact. 95, 1221–1237 (1968).
Raaf, J., Bonner, J.: Properties of Escherichia coli deoxyribonucleoprotein as compared with the deoxyribonucleoprotein of higher organisms. Arch. Biochem. 125, 567–579 (1968).
Seibert, F. B.: Removal of impurities, nucleic acid and polysaccharide from tuberculin protein. J. biol. Chem. 133, 593–604 (1940).
Spiegelmann, S., Aronson, A. J., Fitz-James, P. C.: Isolation and characterization of nuclear bodies from protoplasts of Bacillus megaterium. J. Bact. 75, 10–117 (1958).
Stevens, A.: Incorporation of the adenine ribonucleotide into RNA by cell fractions from E. coli B. Biochem. biophys. Res. Commun. 3, 92–96 (1960).
Szabó, G., Barabás, Gy., Vályi-Nagy, T.: Comparison of Streptomyces griseus strains which produce streptomycin and those which do not. Arch. Mikrobiol. 40, 261–274 (1961).
Szeszák, F., Szabó, G.: Fast sedimenting fractions containing DNA from Streptomyces griseus. Acta microbiol. Acad. Sci. hung. (in press) (1970).
——, Vitális, S.: Isolation of different mycelial fractions from submerged culture of Streptomyces griseus strain by density gradient centrifugation. Zbl. Bakt., II. Abt. 121, 18–40 (1967).
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Szeszák, F., Szabó, G. & Sümegi, J. RNA synthesis on native DNA complexes isolated from Streptomyces griseus and Escherichia coli . Archiv. Mikrobiol. 73, 368–378 (1970). https://doi.org/10.1007/BF00412303
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DOI: https://doi.org/10.1007/BF00412303