Crystallographic observation of nonenzymatic RNA primer extension

ELife
Wen ZhangJack W Szostak

Abstract

The importance of genome replication has inspired detailed crystallographic studies of enzymatic DNA/RNA polymerization. In contrast, the mechanism of nonenzymatic polymerization is less well understood, despite its critical role in the origin of life. Here we report the direct observation of nonenzymatic RNA primer extension through time-resolved crystallography. We soaked crystals of an RNA primer-template-dGMP complex with guanosine-5'-phosphoro-2-aminoimidazolide for increasing times. At early times we see the activated ribonucleotides bound to the template, followed by formation of the imidazolium-bridged dinucleotide intermediate. At later times, we see a new phosphodiester bond forming between the primer and the incoming nucleotide. The intermediate is pre-organized because of the constraints of base-pairing with the template and hydrogen bonding between the imidazole amino group and both flanking phosphates. Our results provide atomic-resolution insight into the mechanism of nonenzymatic primer extension, and set the stage for further structural dissection and optimization of the RNA copying process.

References

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Citations

Jun 1, 2018·ELife·John C Chaput
Mar 26, 2019·Angewandte Chemie·Travis WaltonJack W Szostak
May 20, 2020·Nucleic Acids Research·Daniel DuzdevichJack W Szostak
Jun 20, 2020·Angewandte Chemie·Lijun ZhouJack W Szostak
Nov 9, 2019·ELife·Lijun ZhouJack W Szostak
Feb 27, 2021·Journal of the American Chemical Society·Seohyun Chris KimJack W Szostak
May 13, 2021·Nature Communications·Nicholas ChimJohn C Chaput
Aug 17, 2020·Journal of the American Chemical Society·Stephanie J ZhangJack W Szostak

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Methods Mentioned

BETA
X-ray
NMR

Software Mentioned

Refmac
HKL2000
MestReNova

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