Abstract
Methyltransferase (MTase) transfers a methyl group (-CH3) from the donor S-adenosyl-L-methionine (AdoMet or SAM) to biologically active molecules such as hormones, neurotransmitters, lipids, proteins and nucleic acids. The addition of a methyl group causes a change in the physicochemical properties of the molecules. The mRNA cap structure is essential for cell and virus. Guanine-N7-methyltransferase (N7-MTase) methylates the GpppN cap at the N7 position of guanine, resulting in cap-0 structure (m7GpppN), and Ribose 2'-O-MTase further methylates the first nucleotide of higher eukaryotic cellular and viral mRNAs at the ribose 2'-OH position to form cap-1 (m7GpppNm) structures. Here, we describe a biochemical assay to detect the activities of mRNA capping MTases.
Keywords: Methyltransferase, S-adenosyl-L-methionine, RNA capping, Cap structure
Materials and Reagents
Equipment
Procedure
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Acknowledgments
We thank Dr. Tero Ahola from University of Helsinki for the kindly help and advices for setting up this MTase assay. We thankfully acknowledge the University of Helsinki for providing research facilities and support during our visits. This work was supported by the China NSFC grants (81130083, 31170152 and 81271817).
References
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