Fig. 6: Mutations in GGACA motif of WYMV RNA1 slows the viral infection.
From: N6-methyladenosine RNA modification promotes viral genomic RNA stability and infection
a Schematic diagram of two WYMV R1 mutations. The mutated bases are marked in red. The different colored boxes represent the coding regions of different proteins. The black peak represents m6A peak. b Binding affinity of TaMTB-His(E. coli) with probes 2-5 (right) detected by MST experiment. The mixture with TaMTB protein+ probe 3 was used as a negative control. The solid curve is the fit of the data points to the standard Kd-Fit function. Each binding assay was repeated three times independently, and bars represent SD. Data in (b) are represented as means ± SD. Kd, dissociation constant. The blue sequence represents the m6A motif. c m6A-IP-qPCR assay to detect the m6A level of peak 3 in WYMV-WT or WYMV-mu1/2 infected WT wheat plants and WYMV-mu2-infected TaMTB-OE plants. Values are means ± SD (two-sided t test, n  =  3, P  = 0.0116, P  <  0.0001). **P  <  0.01 *P  <  0.05. ns, no significant. d Schematic diagram of the CP mutation used for mRNA stability assay. The wild type (WT) or mutated (mu) coding region of WYMV CP was cloned into the expression vector driven by the CaMV 35 S promoter. e The mRNA lifetime of WYMV CP. TI: transcription inhibition. Data are represented as means ± SD for 3 biological replicates (two-sided t test, P  = 0.0153, 0.0085). f The accumulation of WYMV RNA1 and RNA2 in WYMV-WT-infected and WYMV-mu2-infected wheat plants were determined by qRT-PCR using CP and P2 gene-specific primers. Values are means ± SD (two-sided t test, n  =  3, P =  0.0192, P  =  0.0021). g Phenotypes in the fourth leaves of the plants inoculated with phosphate buffered saline (MOCK), WYMV-WT or WYMV-mu2, respectively. Source data are provided as a Source Data file.
