Author: Martin, Baptiste; Coutard, Bruno; Guez, Théo; Paesen, Guido C; Canard, Bruno; Debart, Françoise; Vasseur, Jean-Jacques; Grimes, Jonathan M; Decroly, Etienne
Title: The methyltransferase domain of the Sudan ebolavirus L protein specifically targets internal adenosines of RNA substrates, in addition to the cap structure Document date: 2018_9_6
ID: 243u68j8_17
Snippet: To evaluate methyltransferase activities, a radioactive test was set up by mixing 4 M of SUDV MTase+CTD domain with 1 M of purified synthetic RNAs (see all RNAs in Supplementary Table S1), 10 M of SAM and 0.5 M of 3 H-SAM (Perkin Elmer) in an optimized MTase assay buffer (50 mM Tris-HCl pH 8, 10 mM arginine). Reactions were stopped by a 10-fold dilution in water after 3 h at 30 • C. Samples were transferred to DEAE filtermats (Perkin Elmer) usi.....
Document: To evaluate methyltransferase activities, a radioactive test was set up by mixing 4 M of SUDV MTase+CTD domain with 1 M of purified synthetic RNAs (see all RNAs in Supplementary Table S1), 10 M of SAM and 0.5 M of 3 H-SAM (Perkin Elmer) in an optimized MTase assay buffer (50 mM Tris-HCl pH 8, 10 mM arginine). Reactions were stopped by a 10-fold dilution in water after 3 h at 30 • C. Samples were transferred to DEAE filtermats (Perkin Elmer) using a Filtermat Harvester (Packard Instruments). Methyl transfer was then evaluated as described before (26) .
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