PMID: 7028106Sep 15, 1981Paper

Properties of P3 esters of nucleoside triphosphates as substrates for RNA polymerase from Escherichia coli

Biochemistry
W J SmagowiczK H Scheit

Abstract

P3-[(2,4-Dinitrophenyl)amino]ethyl (DNPNHEt) and P3-methyl phosphate esters of nucleoside 5'-triphosphates have been synthesized. Their properties as substrates in the initiation and elongation steps of transcription have been examined by using RNA polymerase from Escherichia coli and poly[d(A-T)] or T7 DNA as templates. It is shown that transcription can be initiated by ATP-EtNHDNP and that 2,4-dinitrophenyl residues are incorporated at the 5' end of the RNA molecules. Steady-state kinetic experiments of abortive initation on promoters A1 and A3 of T7 DNA revealed that ATP-EtNHDNP, ADP-EtNHDNP, and ATP-OCH3 have lower Km values and markedly reduced Vmax values compared to those of ATP. The two classes of esters, NTR-EtNHDNP and NTP-OCH3, were found to differ regarding their utilization as substrates for elongation. Both ATP-OCH3 and UTP-OCH3 are substrates for transcription. However, only the pyrimidine derivatives of NTP-EtNHDNP are elongation substrates which release DNPNHEt-PP upon utilization. This dramatic difference between the purine and pyrimidine derivatives of NTP-EtNHDNP reflects a selective process in the transcriptional complex for purines and pyrimidines.

References

Jan 3, 1977·European Journal of Biochemistry·H R Rackwitz, K H Scheit
Jun 1, 1978·Nucleic Acids Research·C Van Broeckhoven, R De Wachter
Nov 12, 1976·Biochimica Et Biophysica Acta·D C LivingstonD C Ward
Nov 1, 1976·European Journal of Biochemistry·V W Armstrong, F Eckstein
Jun 1, 1976·Ultramicroscopy·W Krakow
Sep 15, 1975·European Journal of Biochemistry·J P DausseP Fromageot
Jun 25, 1973·Journal of Molecular Biology·E G Minkley, D Pribnow
Feb 1, 1970·Hoppe-Seyler's Zeitschrift für physiologische Chemie·W ZilligH J Halbwachs
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Apr 27, 1971·Biochemistry·J L DarlixE Reich
Jul 13, 1972·Biochimica Et Biophysica Acta·R S GoodyR H Schirmer

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Citations

Aug 1, 1982·European Journal of Biochemistry·K H Scheit, H J Linke

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