Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase

Abstract RNA-based drugs are an emerging class of therapeutics combining the immense potential of DNA gene-therapy with the absence of genome integration-associated risks. While the synthesis of such molecules is feasible, large scale in vitro production of humanised mRNA remains a biochemical and e...

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Autores principales:	Pierre-Yves Colin, Paul A. Dalby
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2f9517bf037f477d8cdc334243cc1253
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spelling	oai:doaj.org-article:2f9517bf037f477d8cdc334243cc12532021-12-02T13:58:10ZFunctional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase10.1038/s41598-020-79026-22045-2322https://doaj.org/article/2f9517bf037f477d8cdc334243cc12532020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79026-2https://doaj.org/toc/2045-2322Abstract RNA-based drugs are an emerging class of therapeutics combining the immense potential of DNA gene-therapy with the absence of genome integration-associated risks. While the synthesis of such molecules is feasible, large scale in vitro production of humanised mRNA remains a biochemical and economical challenge. Human mRNAs possess two post-transcriptional modifications at their 5′ end: an inverted methylated guanosine and a unique 2′O-methylation on the ribose of the penultimate nucleotide. One strategy to precisely methylate the 2′ oxygen is to use viral mRNA methyltransferases that have evolved to escape the host’s cell immunity response following virus infection. However, these enzymes are ill-adapted to industrial processes and suffer from low turnovers. We have investigated the effects of homologous and orthologous active-site mutations on both stability and transferase activity, and identified new functional motifs in the interaction network surrounding the catalytic lysine. Our findings suggest that despite their low catalytic efficiency, the active-sites of viral mRNA methyltransferases have low mutational plasticity, while mutations in a defined third shell around the active site have strong effects on folding, stability and activity in the variant enzymes, mostly via network-mediated effects.Pierre-Yves ColinPaul A. DalbyNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-13 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Pierre-Yves Colin Paul A. Dalby Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase
description	Abstract RNA-based drugs are an emerging class of therapeutics combining the immense potential of DNA gene-therapy with the absence of genome integration-associated risks. While the synthesis of such molecules is feasible, large scale in vitro production of humanised mRNA remains a biochemical and economical challenge. Human mRNAs possess two post-transcriptional modifications at their 5′ end: an inverted methylated guanosine and a unique 2′O-methylation on the ribose of the penultimate nucleotide. One strategy to precisely methylate the 2′ oxygen is to use viral mRNA methyltransferases that have evolved to escape the host’s cell immunity response following virus infection. However, these enzymes are ill-adapted to industrial processes and suffer from low turnovers. We have investigated the effects of homologous and orthologous active-site mutations on both stability and transferase activity, and identified new functional motifs in the interaction network surrounding the catalytic lysine. Our findings suggest that despite their low catalytic efficiency, the active-sites of viral mRNA methyltransferases have low mutational plasticity, while mutations in a defined third shell around the active site have strong effects on folding, stability and activity in the variant enzymes, mostly via network-mediated effects.
format	article
author	Pierre-Yves Colin Paul A. Dalby
author_facet	Pierre-Yves Colin Paul A. Dalby
author_sort	Pierre-Yves Colin
title	Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase
title_short	Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase
title_full	Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase
title_fullStr	Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase
title_full_unstemmed	Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase
title_sort	functional and computational identification of a rescue mutation near the active site of an mrna methyltransferase
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/2f9517bf037f477d8cdc334243cc1253
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Functional and computational identification of a rescue mutation near the active site of an mRNA methyltransferase

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