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Dinucleoside Polyphosphates as RNA Building Blocks with Pairing Ability in Transcription Initiation
R. Benoni, M. Culka, O. Hudeček, L. Gahurova, H. Cahová
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Bacteriophage T7 enzymology MeSH
- Dinucleoside Phosphates genetics metabolism MeSH
- DNA-Directed RNA Polymerases genetics metabolism MeSH
- DNA metabolism MeSH
- Transcription Initiation, Genetic * MeSH
- Base Pairing MeSH
- RNA Caps genetics MeSH
- RNA genetics metabolism MeSH
- Molecular Dynamics Simulation MeSH
- Protein Binding MeSH
- Viral Proteins genetics metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Dinucleoside polyphosphates (NpnNs) were discovered 50 years ago in all cells. They are often called alarmones, even though the molecular target of the alarm has not yet been identified. Recently, we showed that they serve as noncanonical initiating nucleotides (NCINs) and fulfill the role of 5' RNA caps in Escherichia coli. Here, we present molecular insight into their ability to be used as NCINs by T7 RNA polymerase in the initiation phase of transcription. In general, we observed NpnNs to be equally good substrates as canonical nucleotides for T7 RNA polymerase. Surprisingly, the incorporation of ApnGs boosts the production of RNA 10-fold. This behavior is due to the pairing ability of both purine moieties with the -1 and +1 positions of the antisense DNA strand. Molecular dynamic simulations revealed noncanonical pairing of adenosine with the thymine of the DNA.
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