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Electron microscopic analysis of in vitro transcriptional complexes: Mapping of promoters of the coliphage T5 genome

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Summary

Transcriptional complexes formed in vitro with coliphage T5 DNA as template were analyzed by electron microscopy and the number and location of starting sites utilized by E. coli RNA polymerase were determined. Of the 40 promoters characterized in this way, 6 map in the two terminal “pre-early” regions, 29 in the “early” and 5 in the “late” region. The direction of transcription within the different regions determined in this study agrees with earlier findings derived from RNA synthesized in vivo.

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References

  • Alberts, B.M., Frey, L.: T 4 bacteriophage gene 32: A structural protein in the replication and recombination of DNA. Nature 227, 1313–1318 (1970)

    Google Scholar 

  • Bordier, C., Dubochet, J.: Electron microscope localisation of the binding sites of E. coli RNA polymerase in the early promoter region of T7 DNA. Europ. J. Biochem. 44, 617–624 (1974)

    Google Scholar 

  • Bourguignon, G.J., Sweeney, T. K., Delius, H.: Multiple origins and circular structures in replicating T5 bacteriophage DNA. J. Virol. 18, 245–259 (1976)

    Google Scholar 

  • Brunschede, H., Bremer H.: Synthesis of phage and hostspecific RNA in E. coli infected with a fragment of bacteriophage T5. Virology 75, 355–367 (1976)

    Google Scholar 

  • Bujard, H., Hendrickson, H.E.: Structure and function of the genome of coliphage T5. I. The physical structure of the chromosome of T5+. Europ. J. Biochem. 33, 517–528 (1973)

    Google Scholar 

  • Davis, R., Hyman: Physical locations of the in vitro RNA initiation site and termination sites on T7 M DNA. Cold Spr. Harb. Symp. quant. Biol. 35, 269–281 (1971)

    Google Scholar 

  • Delius, H., Westphal, H., Axelrod, N.: Length measurement of RNA synthesized in vitro by Escherichia coli RNA polymerase, J. molec. Biol. 74, 677–687 (1973)

    Google Scholar 

  • Dresel, H.A., Diplomarbeit, Universität Heidelberg, 1976

  • Dresel, H.A., Gabain, A.v., Hirth, K.P., Fuchs, E., Bujard, H.: The pattern of RNA synthesis and its interdependence with protein synthesis during the development of coliphage T5. (In preparation) (1978a)

  • Dunn, J.J., Studier, F.W.: T7 early RNAs are generated by site-specific cleavages. Proc. nat. Acad. Sci. (Wash.) 70, 1559–1563 (1973)

    Google Scholar 

  • Gabain, A.v., Bujard, H.: Interaction of E. coli RNA polymerase with promoters of coliphage T5. The rates of complex formation and decay and their correlation with in vitro and in vivo transcriptional activity. Molec. gen. Genet. 157, 301–311 (1977)

    Google Scholar 

  • Gabain, A.v., Bujard, H.: The binding of E. coli RNA polymerase to promoters of various E. coli systems. Proc. nat. Acad. Sci. (Wash). (Submitted) (1978)

  • Gabain, A.v., Hayward, G.S., Bujard, H.: Physical mapping of the Hind III, Eco RI, Sal and Sma restriction endonuclease cleavage fragments from bacteriophage T5 DNA. Molec. gen. Genet. 143, 279–290 (1976)

    Google Scholar 

  • Hendrickson, H.E., Bujard, H.: Structure and function of the genome of coliphage T5. 2. Regions of transcription of the chromosomes. Europ. J. Biochem. 33, 529–534 (1973)

    Google Scholar 

  • Knopf, K.W., Bujard, H.: Structure and function of the genome of coliphage T5: 3. Transcription in vitro of the “nicked” and “nickfree” T5+ DNA. Europ. J. Biochem. 53, 371–385 (1975)

    Google Scholar 

  • Koller, Th., Sogo, J., Bujard, H.: An electron microscope method for studying nucleic acid-protein complexes. Visualization of RNA polymerase bound to the DNA of bateriophage T7 and T3. Biopolymers 13, 995–1009 (1974)

    Google Scholar 

  • Lanni, Y.T.: Invasion by bacteriophage T5. II. Dissociation of calcium-independent and calcium-dependent processes. Virology 10, 514–529 (1960)

    Google Scholar 

  • McCorquodale, D.J.: The T-odd bacteriophages. CRC Crit. Rev. Microbiol. 4, 101–159 (1975)

    Google Scholar 

  • Pispa, J.P., Buchanan, J.M.: Synthesis of bacteriophage specific RNA in vitro. Biochem. biophys. Acta (Amst.) 247, 181–184 (1971)

    Google Scholar 

  • Saigo, K.: Denaturation mapping and chromosome structure in bacteriophage T5. Virology 68, 166–172 (1975)

    Google Scholar 

  • Saigo, K.: A new bacteriophage T5 particle with a DNA lacking the terminal repetition at the “right hand” end. J. molec. Biol. 107, 369–378 (1976)

    Google Scholar 

  • Stüber, D., Bujard, H.: Electron microscopy of DNA: Determination of absolute molecular weights and linear density. Molec. gen. Genet. 154, 299–303 (1977)

    Google Scholar 

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Communicated by H.G. Wittmann

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Stüber, D., Delius, H. & Bujard, H. Electron microscopic analysis of in vitro transcriptional complexes: Mapping of promoters of the coliphage T5 genome. Molec. Gen. Genet. 166, 141–149 (1978). https://doi.org/10.1007/BF00285916

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  • DOI: https://doi.org/10.1007/BF00285916

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