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

分子拥挤效应通过碱基堆积作用诱导RNA聚合酶进行引物延伸,这种作用超越了沃森-克里克规则。

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作者:Shuntaro Takahashi,Hiromichi Okura,Pallavi Chilka,Saptarshi Ghosh,Naoki Sugimoto

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.

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