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Electron microscopic mapping and sequence analysis of the terminator for the early message of E. coli phage T7

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Summary

The terminator position of T7 early messenger RNA was determined by electron microscopic measurements. The end of the RNA was mapped at a position 18.9% from the left end of T7 DNA, and 145±25 nucleotides from the right end of the Hpa I fragment Q. The sequence of the Hpa I Q fragment was determined around this position, and a terminator-like structure was detected in position 193 to 169 from the right end of fragment Q.

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References

  • Aaij, C., Borst, P.: The gel electrophoresis of DNA. Biochem. Biophys. Acta 269, 192–200 (1972)

    Google Scholar 

  • Adhya, S., Gottesman, M.: Control of transcription termination. Annu. Rev. Biochem. 47, 967–996 (1978)

    Google Scholar 

  • Blin, N., Gabain, A. v., Bujard, H.: Isolation of large molecular weight DNA from agarose gels for further digestion by restriction enzymes. FEBS Lett. 53, 84–86 (1975)

    Google Scholar 

  • Bordier, C., Dubochet, J.: Electron microscopic localization of the binding sites of Escherichia coli RNA polymerase in the early promoter region of T7 DNA. Eur. J. Biochem. 44, 617–624 (1974)

    Google Scholar 

  • Chow, L.T., Roberts, J.M., Lewis, J.B., Broker, T.R.: A map of cytoplasmic RNA transcripts from lytic adenovirus type 2, determined by electron microscopy of RNA: DNA hybrids. Cell 11, 819–836 (1977)

    Google Scholar 

  • Delius, H., Mantell, N.J., Alberts, B.: Characterization by electron microscopy of the complex formed between T4 bacteriophage gene 32-protein and DNA. J. Mol. biol. 67, 341–350 (1972)

    Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  • Dunn, J.J., Studier, F.W.: T7 early RNAs and E. coli ribosomal RNAs are cut from large precursor RNAs in vivo by ribonuclease III. Proc. Natl. Acad. Sci. U.S.A. 70, 3296–3300 (1973b)

    Google Scholar 

  • Griffith, J.D.: DNA structure: Evidence from electron microscopy. Science 201, 525–527 (1978)

    Google Scholar 

  • Hayward, G.S.: Gel electrophoretic separation of the complementary strands of bacteriophage DNA. Virology 49, 342–344 (1972)

    Google Scholar 

  • Hobom, G., Grosschedl, R., Lusky, M., Scherer, G., Schwarz, E., Koessel, H.: Functional analysis of the replicator structure of lambdoid bacteriophage DNAs. Cold Spring Harbor Symp. Quant. Biol. 43, 165–178 (1978)

    Google Scholar 

  • Hyman, R.W.: Physical mapping of T7 messenger RNA. J. Mol. Biol. 61, 369–376 (1971)

    Google Scholar 

  • Koller, B., Delius, H., Buenemann, H., Mueller, W.: The isolation of DNA from agarose gels by electrophoretic elution onto malachite green-polyacrylamide columns. Gene 4, 227–239 (1978b)

    Google Scholar 

  • Koller, Th., Kuebler, O., Portmann, R., Sogo, J.M.: High resolution mapping of specific binding sites of Escherichia coli RNA polymerase on the DNA of bacteriophage T7. J. Mol. Biol. 120, 121–131 (1978a)

    Google Scholar 

  • Korobko, V.G., Chuvpilo, S.A., Grachev, S.A., Kolossov, M. N., Grachev, M.A., Zaychikov, E.F., Pletryov, A. G.: Nucleotide-sequence of the A1-A3 promoter region of phage T7. Bioorg. Khim. 4, 1692–1694 (1978)

    Google Scholar 

  • Maxam, A.M., Gilbert, W.: A new method for sequencing DNA. Proc. Natl. Acad. Sci. U.S.A. 74, 560–564 (1977)

    Google Scholar 

  • Maxam, A.M., Gilbert, W.: Sequencing endlabeled DNA with base specific chemical cleavages. In: Methods in Enzymology (Grossman, L., Moldave, K., eds.), vol. 65. New York: Academic Press (in press)

  • Maitra, U., Lockwood, A.H., Dubnoff, J.S., Guha, A.: Termination, release, and reinitiation of RNA chains from DNA templates by Escherichia coli RNA polymerase. Cold Spring Harbor Symp. Quant. Biol. 35, 143–156 (1970)

    Google Scholar 

  • McDonell, M.W., Simon, M.N., Studier, F.W.: Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J. Mol. Biol. 110, 119–146 (1977)

    Google Scholar 

  • Millette, R.L., Trotter, C.D., Herrlich, P., Schweiger, M.: In vitro synthesis, termination, and release of active messenger RNA. Cold Spring Harbor Symp. Quant. Biol. 35, 135–142 (1970)

    Google Scholar 

  • Peacock, C.A., Dingman, C.W.: Resolution of multiple ribonucleic acid species by polyacrylamide gel electrophoresis. Biochemistry 6, 1818–1827 (1967)

    Google Scholar 

  • Pettersson, U., Mulder, C., Delius, H., Sharp, P.A.: Cleavage of adenovirus type 2 DNA into six unique fragments by endonuclease R. RI. Proc. Natl. Acad. Sci. U.S.A. 70, 200–204 (1973)

    Google Scholar 

  • Portmann, R., Koller, Th.: The divalent cation method for protein free spreading of nucleic acid molecules. Sixth European Congr. Electron Microscopy Jerusalem, vol. 2, pp. 546–548. Tal International Publishing Company 1976

  • Portmann, R., Sogo, J.M., Koller, Th., Zillig, W.: Binding sites of E. coli RNA polymerase on T7 DNA as determined by electron microscopy. FEBS Lett. 45, 64–67 (1974)

    Google Scholar 

  • Pribnow, D.: Bacteriophage T7 early promoters: Nucleotide sequences of two RNA polymerase binding sites. J. Mol. Biol. 99, 419–443 (1975)

    Google Scholar 

  • Rosenberg, M., Kramer, R.A., Steitz, J.A.: T7 early messenger RNAs are the direct products of ribonuclease III cleavage. J. Mol. Biol. 89, 777–782 (1974)

    Google Scholar 

  • Sanger, F., Coulson, A.R.: The use of thin acrylamide gels for DNA sequencing. FEBS Lett. 87, 107–110 (1978)

    Google Scholar 

  • Siebenlist, U.: Nucleotide sequence of the three major early promoters of bacteriophage T7. Nucleic Acids Res. 6, 1895–1907 (1979)

    Google Scholar 

  • Sutcliffe, J.G.: Complete nucleotide sequence of the Escherichia coli plasmid pBR322. Cold Spring Harbor Symp. Quant. Biol. 43, 77–90 (1978)

    Google Scholar 

  • Tabak, H.F., Flavell, R.A.: A method for the recovery of DNA from agarose gels. Nucleic Acids Res. 5, 2321–2332 (1978)

    Google Scholar 

  • Williams, R.C.: Use of polylysine for adsorption of nucleic acids and enzymes to electron microscope specimen films. Proc. Natl. Acad. Sci. U.S.A. 74, 2311–2315 (1977)

    Google Scholar 

  • Wu, M., Davidson, N.: Use of gene 32-protein staining of single-stranded polynucleotides for gene mapping by electron microscopy; application to the ϕ80 d3 ilv su +7 system. Proc. Natl. Acad. Sci. U.S.A. 72, 4506–4510 (1975)

    Google Scholar 

  • Zollinger, M., Guertin, M., Mamet-Bratley, M.D.: A new electron microscopic method for studying protein-nucleic acid interactions. Anal. Biochem. 82, 196–203 (1977)

    Google Scholar 

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Communicated by H. Saedler

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Priess, H., Koller, B., Hess, B. et al. Electron microscopic mapping and sequence analysis of the terminator for the early message of E. coli phage T7. Molec. Gen. Genet. 178, 27–34 (1980). https://doi.org/10.1007/BF00267209

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  • DOI: https://doi.org/10.1007/BF00267209

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