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Issue 55, 2020
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Molecular crowding induces primer extension by RNA polymerase through base stacking beyond Watson–Crick rules

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Abstract

The polymerisation of nucleic acids is essential for copying genetic information correctly to the next generations, whereas mispolymerisation could promote genetic diversity. It is possible that in the prebiotic era, polymerases might have used mispolymerisation to accelerate the diversification of genetic information. Even in the current era, polymerases of RNA viruses frequently cause mutations. In this study, primer extension under different molecular crowding conditions was measured using T7 RNA polymerase as a model for the reaction in the prebiotic world. Interestingly, molecular crowding using 20 wt% poly(ethylene glycol) 2000 preferentially promoted the primer extensions with ATP and GTP by T7 RNA polymerase, regardless of Watson–Crick base-pairing rules. This indicates that molecular crowding decreases the dielectric constants in solution, resulting in enhancement of stacking interactions between the primer and an incorporated nucleotide. These findings suggest that molecular crowding could accelerate genetic diversity in the prebiotic world and may promote transcription error of RNA viruses in the current era.

Graphical abstract: Molecular crowding induces primer extension by RNA polymerase through base stacking beyond Watson–Crick rules

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Supplementary files

Article information


Submitted
27 Jul 2020
Accepted
27 Aug 2020
First published
07 Sep 2020

This article is Open Access

RSC Adv., 2020,10, 33052-33058
Article type
Paper

Molecular crowding induces primer extension by RNA polymerase through base stacking beyond Watson–Crick rules

S. Takahashi, H. Okura, P. Chilka, S. Ghosh and N. Sugimoto, RSC Adv., 2020, 10, 33052
DOI: 10.1039/D0RA06502A

This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence. Material from this article can be used in other publications provided that the correct acknowledgement is given with the reproduced material and it is not used for commercial purposes.

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    [Original citation] - Published by The Royal Society of Chemistry (RSC) on behalf of the European Society for Photobiology, the European Photochemistry Association, and RSC.
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    [Original citation] - Published by The Royal Society of Chemistry.

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