Fig. 4: TaMTb is a member of writers of m6A in wheat.
From: N6-methyladenosine RNA modification promotes viral genomic RNA stability and infection
a Phylogenetic and conserved domain analysis of TaMTB from difference species. The MTB protein sequences were obtained from Homo sapiens (hs), Mus musculus (mrce), Zea mays (Zm), Sorghum bicolor (Sb), Oryza sativa (Os), Vitis vinifera (Vv), Gossypium hirsutum (Gh), T. aestivum (Ta), Arabidopsis thaliana (At), Brassica rapa (Br). TaMTB is highlighted in the phylogenetic tree using a red circle. The phylogenetic tree was constructed by the neighbor-joining method in MEGA 7.0 software, with bootstrap values of 1000. The percentage on phylogenetic tree nodes indicated that the associated taxa clustered together in the bootstrap test. The branch lengths represent evolutionary distances, which are calculated using p-distance method. b In vitro methylation assay of TaMTB by detection with dot blot. The part marked in red represents the m6A motif. The RNA oligo with m6A modification (listed in Supplementary Data 3) was used as positive control. IP-GFP-Flag and His protein was used as negative controls. IP-TaMTB-Flag and IP-GFP-Flag were purified from 35 S:TaMTB-Flag and 35 S:GFP-Flag transfected wheat protoplasts. TaMTB-His (E. coli) was purified from E. coli. TaMTB-His (Sf9) was purified from Sf9 cells. c, d, e Real-time fluorescence amplification curves and bar plot of the threshold cycle (CT) of qPCR showing SELECT results for detecting the m6A site in RNA oligo with IP-TaMTB-Flag, TaMTB-His (E. coli) and TaMTB-His (Sf9) treatment. Rn is the raw fluorescence for the associated well normalized to the fluorescence of the passive reference dye (ROX). Values are means ± SD (two-sided t test, n  =  3, (c) P  <  0.0001). **P  <  0.01. ns, not significant. The red or yellow marked A represent the base that are possible to be m6A methylated. Source data are provided as a Source Data file.
