Phosphonate analogues of dinucleotides as substrates for DNA-dependent RNA polymerase from Escherichia coli in primed abortive initiation reaction
Language English Country Netherlands Media print
Document type Journal Article
PubMed
2489057
DOI
10.1016/0141-8130(89)90037-8
PII: 0141-8130(89)90037-8
Knihovny.cz E-resources
- MeSH
- Dinucleoside Phosphates metabolism MeSH
- DNA-Directed RNA Polymerases metabolism MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Escherichia coli enzymology MeSH
- Transcription, Genetic MeSH
- Kinetics MeSH
- Organophosphonates MeSH
- Poly dA-dT metabolism MeSH
- Substrate Specificity MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Dinucleoside Phosphates MeSH
- DNA-Directed RNA Polymerases MeSH
- Organophosphonates MeSH
- Poly dA-dT MeSH
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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