Phosphonate analogues of dinucleotides as substrates for DNA-dependent RNA polymerase from Escherichia coli in primed abortive initiation reaction

Ales Cvekl, Květa Horská, Karel Sěbesta, Ivan Rosenberg, Antoním Holý

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Dinucleotides (3′-5′)-ApU and UpA and their 3′-O-phosphonylmethyl and 5′-O-phosphonylmethyl analogues were studied as substrates in the primed abortive synthesis catalysed by Escherichia coli DNA-dependent RNA polymerase on poly[d(A-T)] template. All phosphonate analogues of dinucleotides containing the anomalous sugar-phosphate backbone are substrates for the holoenzyme as verified by RNase A and RNase T2 digestion of the trinucleotide analogues obtained. The finding that phosphonate dinucleotides act as primers for transcription indicates that steric requirements at the initiation site are not as specific as previously supposed. Analysis of kinetic constants of ordered bibi reaction Kia, KmA, KmB and Vmax suggests that the instability of short RNA-DNA hybrids contributes to the abortive release of trinucleotides formed.

Original languageEnglish (US)
Pages (from-to)33-38
Number of pages6
JournalInternational Journal of Biological Macromolecules
Volume11
Issue number1
DOIs
StatePublished - 1989
Externally publishedYes

Fingerprint

Organophosphonates
ribonuclease T(2)
DNA-Directed RNA Polymerases
Escherichia coli
Sugar Phosphates
Pancreatic Ribonuclease
Holoenzymes
RNA Stability
Substrates
Transcription
Digestion
RNA
Kinetics
DNA
2'-deoxythymidylyl-(3'-5')-2'-deoxyadenosine
poly A-T

Keywords

  • phosphonate dinucleotide
  • RNA polymerase
  • transcription

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Phosphonate analogues of dinucleotides as substrates for DNA-dependent RNA polymerase from Escherichia coli in primed abortive initiation reaction. / Cvekl, Ales; Horská, Květa; Sěbesta, Karel; Rosenberg, Ivan; Holý, Antoním.

In: International Journal of Biological Macromolecules, Vol. 11, No. 1, 1989, p. 33-38.

Research output: Contribution to journalArticle

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AU - Horská, Květa

AU - Sěbesta, Karel

AU - Rosenberg, Ivan

AU - Holý, Antoním

PY - 1989

Y1 - 1989

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AB - Dinucleotides (3′-5′)-ApU and UpA and their 3′-O-phosphonylmethyl and 5′-O-phosphonylmethyl analogues were studied as substrates in the primed abortive synthesis catalysed by Escherichia coli DNA-dependent RNA polymerase on poly[d(A-T)] template. All phosphonate analogues of dinucleotides containing the anomalous sugar-phosphate backbone are substrates for the holoenzyme as verified by RNase A and RNase T2 digestion of the trinucleotide analogues obtained. The finding that phosphonate dinucleotides act as primers for transcription indicates that steric requirements at the initiation site are not as specific as previously supposed. Analysis of kinetic constants of ordered bibi reaction Kia, KmA, KmB and Vmax suggests that the instability of short RNA-DNA hybrids contributes to the abortive release of trinucleotides formed.

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