Summary
The biological significance of the low level of symmetric and non-specific RNA synthesis catalyzed by the core RNA polymerase devoid of the sigma factor has been analyzed. Shearing of DNA's including T4 DNA markedly increased the template activities with the core enzyme but not with the holoenzyme. This finding suggests that RNA synthesis by the core enzyme increases concomittantly with the production of termini in DNA. Double-stranded circular DNA's such as λdv and fd-RFI were found to be inactive as templates for the core enzyme, but were made active by introduction of single-strand nicks with deoxyribonuclease. In contrast, single-stranded circular DNA (ϕX 174) served as a good template for RNA synthesis by the core RNA polymerase. These findings suggest that the sigma factor may activate double-stranded DNA at the promotor sites by creating proper initiation points for RNA synthesis. Partial separation of duplex DNA into single-stranded forms at the promotor sites could be one of the processes in the reaction catalyzed by the holoenzyme containing the sigma factor.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Anthony, D. D., Zeszotek, E., Goldthwait, D. A.: Studies with RNA polymerase, II. Kinetic aspects of initiation and polymerization. Biochemistry (Wash.) 8, 246–256 (1969).
Bautz, E. K., Bautz, F. A., Dunn, J. J.: E. coli sigma factor: a positive control element in phage T4 development. Nature (Lond.) 223, 1022–1034 (1969).
Burgess, R. R., Travers, A. A., Dunn, G. G., Bautz, E. K. F.: Factor stimulating transcription by RNA polymerase. Nature (Lond.) 221, 43.46 (1969).
Hershey, A. D., Burgi, E.: Molecular homogeneity of the deoxyribonucleic acid of phage T2. J. molec. Biol. 2, 143–152 (1960).
Ishihama, A., Hurwitz, J.: The role of deoxyribonucleic acid in ribonucleic acid synthesis, XVII. The multiple active sites of E. coli RNA polymerase. J. biol. Chem. 244, 6680–6695 (1969).
—, Kameyama, T.: The molecular mechanism of the enzymic reaction in RNA synthesis. Biochim. biophys. Acta (Amst.) 138, 480–498 (1967).
Kameyama, T., Kitano, Y., Kawakami, H., Iida, Y., Murakami, S., Tanaka, Y., Ishihama, A.: The molecular structures and functions of RNA polymerase of Escherichia coli. Nucleic acid metabolism, cell differentiation and cancer growth, ed. by E. V. Cowdry and S. Seno, p. 29–46. Oxford-New York: Pergamon Press 1969.
Kay, E. R. M., Simmons, N. S., Dounce, A. L.: An improved preparation of sodium deoxyribonucleate. J. Amer. chem. Soc. 74, 1724–1726 (1952).
Krakow, J. S., Daley, K., Karstadt, M.: Azotobacter vinelandii RNA polymerase, VII. Enzyme transitions during unprimed r[1-C] synthesis. Proc. nat. Acad. Sci. (Wash.) 62, 432–437 (1969).
Matsubara, K., Kaiser, A. D.: λdv, an autonomously replicating DNA fragment. Cold Spr. Harb. Symp. quant. Biol. 33, 769–776 (1968).
Sinsheimer, R. L.: Isolation, preparation, and characterization of natural and synthetic nucleic acids, sect. B-3. Single-stranded DNA. Procedures in Nucleic Acid Research, ed. by G. L. Cantoni, and D. R. Davies, p. 569–576. New York: Haper & Row, Publ. 1966.
Sugiura, M., Okamoto, T., Takanami, M.: RNA polymerase sigma-factor and the selection of initiation site. Nature (Lond.) 225, 598–600 (1970).
Summers, W. C.: Control of template specificity of E. coli RNA polymerase by a phage-coded protein. Nature (Lond.) 223, 1111–1113 (1969).
Taketo, A.: Studies on the infections DNA from bacterial virus ΦX 174 and the virus-infected cells. J. Biochem. (Tokyo) 54, 520–529 (1963).
Travers, A. A.: Bacteriophage sigma factor for RNA polymerase. Nature (Lond.) 223, 1107–1110 (1969).
— Burgess, R. R.: Cyclic re-use of the RNA polymerase sigma factor. Nature (Lond.) 222, 537–540 (1969).
Vogt, V.: Breaks in DNA stimulate transcription by core RNA polymerase. Nature (Lond.) 223, 854–855 (1969).
Author information
Authors and Affiliations
Additional information
Communicated by Jun-ichi Tomizawa
Rights and permissions
About this article
Cite this article
Ishihama, A., Murakami, S., Fukuda, R. et al. The nature of initiation sites on DNA for the core RNA polymerase. Molec. Gen. Genet. 111, 66–76 (1971). https://doi.org/10.1007/BF00286555
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00286555