Enzymatic RNA Production from NTPs Synthesized from Nucleosides and Trimetaphosphate**
| Autor: | Fabio Chizzolini, Luiz F. M. Passalacqua, Andrej Lupták, Alexandra D. Kent |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Phosphorous Acids
010402 general chemistry 01 natural sciences Biochemistry Viral Proteins Reaction rate constant RNTP medicine T7 RNA polymerase RNA Catalytic Molecular Biology Polymerase Molecular Structure biology 010405 organic chemistry Chemistry Organic Chemistry Ribozyme RNA DNA-Directed RNA Polymerases Ribonucleotides Ribonucleoside Combinatorial chemistry 0104 chemical sciences biology.protein Molecular Medicine Nucleoside medicine.drug |
| Zdroj: | ChemBioChem. 22:2098-2101 |
| ISSN: | 1439-7633 1439-4227 |
| DOI: | 10.1002/cbic.202100085 |
| Popis: | A mechanism of nucleoside triphosphorylation would have been critical in an evolving "RNA world" to provide high-energy substrates for reactions such as RNA polymerization. However, synthetic approaches to produce ribonucleoside triphosphoates (rNTPs) have suffered from conditions such as high temperatures or high pH that lead to increased RNA degradation, as well as substrate production that cannot sustain replication. Previous reports have demonstrated that cyclic trimetaphosphate (cTmp) can react with nucleosides to form rNTPs under prebiotically-relevant conditions, but their reaction rates were unknown and the influence of reaction conditions not well-characterized. Here we established a sensitive assay that allowed for the determination of second-order rate constants for all four rNTPs, ranging from 1.7 x 10 -6 to 6.5 x 10 -6 M -1 s -1 . The ATP reaction shows a linear dependence on pH and Mg 2+ , and an enthalpy of activation of 88 ± 4 kJ/mol. At millimolar nucleoside and cTmp concentrations, the rNTP production rate is sufficient to facilitate RNA synthesis by both T7 RNA polymerase and a polymerase ribozyme. We suggest that the optimized reaction of cTmp with nucleosides may provide a viable connection between prebiotic nucleotide synthesis and RNA replication. |
| Databáze: | OpenAIRE |
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