Initiation of RNA molecules by purified Escherichia coli RNA polymerase (original) (raw)
Summary
The kinesties of appearance of, and the distribution among, the four bases of the chain-initial nucleotides (5′-termini) of RNA chains synthesized in vitro under various conditions have been investigated. The results of this study have shown that when native T4 bacteriophage DNA is the template for the RNA polymerase, most chains start with a purine nucleotide. The ratio of ATP termini to GTP termini is independent of the reaction time and of the template/enzyme ratio in the reaction mixture. A similar preferential purine initiation was observed when denatured T4 is the template, but the ratio of ATP to GTP termini is reduced. All 5′-termini of poly-AU chains synthesized on poly-dAT templates are ATP.
The determination of the kinetics of initiation of RNA chains has allowed direct confirmation of some conclusions which had been inferred previously from sedimentation analyses of the RNA product. (1) Most RNA chains are initiated during a short period at the outset of the reaction. (2) Low concentrations of native DNA templates limit the number of RNA chains synthesized, not the rate of RNA chain growth. (3) The maximum number of RNA molecules which can be synthesized on denatured DNA templates is severalfold larger than the maximum number which can be synthesized on the same weight of native DNA.
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References
- Ames, B. N., R. F. Goldberger, P. E. Hartman, R. G. Martin, and J. R. Roth: The histidine operon. In: Regulation of nucleic acid and protein biosynthesis, p. 272–287 (V. V. Koningsberger and L. Bosch, eds.). Amsterdam: Elsevier 1967.
Google Scholar - Bremer, H.: Chain growth rate and length of enzymatically synthesized RNA molecules. Molec. gen. Genetics 99, 362–371 (1967).
Google Scholar - —, and M. W. Konrad: A complex of enzymatically synthesized RNA and template DNA. Proc. nat. Acad. Sci. (Wash.) 51, 801–808 (1964).
Google Scholar - ——, and R. Bruner: Capacity of T4 DNA to serve as template for purified Escherichia coli RNA polymerase. J. molec. Biol. 16, 104–117 (1966a).
Google Scholar - —— K. Gaines, and G. S. Stent: Direction of chain growth in enzymic RNA synthesis. J. molec. Biol. 13, 540–533 (1965).
Google Scholar - — C. Yegian, and M. Konrad: Inactivation of purified Escherichia coli RNA polymerase by transfer RNA. J. molec. Biol. 16, 94–103 (1966b).
Google Scholar - Chamberlin, M., and P. Berg: Deoxyribonucleic acid-directed synthesis of ribonucleic acid by an enzyme from Escherichia coli. Proc. nat. Acad. Sci. (Wash.) 48, 81–94 (1962).
Google Scholar - Fox, C. F., R. I. Gumport, and S. B. Weiss: The enzymatic synthesis of ribonucleic acid. V. The interaction of ribonucleic acid polymerase with nucleic acids. J. biol. Chem. 240, 2101–2109 (1965).
Google Scholar - Fuchs, E., W. Zillig, P. H. Hofschneider, and A. Preuss: Preparation and properties of RNA-polymerase particles. J. molec. Biol. 10, 546–550 (1964).
Google Scholar - Geiduschek, E., L. Snyder, A. Colvill, and M. Sarnat: Selective synthesis of T-even bacteriophage messenger in vitro. J. molec. Biol. 19, 541–547 (1966).
Google Scholar - Green, M. H.: Strand selective transcription of T4 DNA in vitro. Proc. nat. Acad. Sci. (Wash.) 52, 1388–1395 (1964).
Google Scholar - Krakow, J. S.: Azotobacter vinelandii ribonucleic acid polymerase. II. Effect of ribonuclease on polymerase activity. J. biol. Chem. 241, 1830–1834 (1966).
Google Scholar - Luria, S. E.: Asymmetric transcription of T4 phage DNA by purified RNA polymerase. Biochem. biophys. Res. Commun. 18, 735–742 (1965).
Google Scholar - Maitra, U., and J. Hurwitz: The role of DNA in RNA synthesis. IX. Nucleoside triphosphate termini in RNA polymerase products. Proc. nat. Acad. Sci. (Wash.) 54, 815–822 (1965).
Google Scholar - Shin, D., and K. Moldave: Effect of ribosomes on the biosynthesis of ribonucleic acid in vitro. J. molec. Biol. 21, 231–245 (1966).
Google Scholar
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Authors and Affiliations
- Virus Laboratory, University of California, 94720, Berkeley, California, USA
Hans Bremer & Robert Bruner - Department of Molecular Biology, University of California, 94720, Berkeley, California, USA
Hans Bremer & Robert Bruner - Division of Biology, Southwest Center for Advanced Studies, Post Office Box 30365, 75230, Dallas, Texas, USA
Hans Bremer
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- Hans Bremer
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Bremer, H., Bruner, R. Initiation of RNA molecules by purified Escherichia coli RNA polymerase.Molec. Gen. Genet. 101, 6–16 (1968). https://doi.org/10.1007/BF00434807
- Received: 11 August 1967
- Issue Date: March 1968
- DOI: https://doi.org/10.1007/BF00434807