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bioRxiv · 10.1101/2020.12.22.424046

Precise Regulation and Site-specifically Covalent Labeling of NSUN2 Enable by Genetic Encoding Expansion

Wang, R.·Zhao, J.·Hu, H.·Wang, S.·Wang, L.
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Abstract

RNA plays a critical role in gene expression regulation, cell migration, differentiation, cell death in living organism. 5-Methylcytosine is a post transcriptional RNA modification identified across wide ranges of RNA species including message RNAs. It is reported the addition of m5C to RNA cytosines is enabled by use of NSUN family enzyme, NSUN2 is identified as a critical RNA methyltransferase for adding m5C to mRNA. We demonstrated here that natural lysines modified with special groups were synthesized via chemical synthesis. Through two rounds of positive screening and one round of negative screening, MbPylRS-tRNAcua unnatural lysine substitution system which can specifically recognize lysine with defined group was evaluated and identified. Non-natural lysine substitution at C271 of NSUN2 active site and subsequently fluorescent labeling was realized via so-called click reaction. The function of NSUN2 mutant and its upregulated CDK1 gene and its effect on cell proliferation were also evaluated.

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Wang, R., Zhao, J., Hu, H., Wang, S., Wang, L.. 2020-12-22. Precise Regulation and Site-specifically Covalent Labeling of NSUN2 Enable by Genetic Encoding Expansion. https://doi.org/10.1101/2020.12.22.424046

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