Fig. 1
Methylation of VTRNA1.1 by NSUN2 determines the biogenesis of svRNA4. a–c Correlation of site-specific methylation (m5C) levels (a, b) and methylation level at all covered cytosines in VTRNA1.1 (c) in NSUN2-/- cells infected with the empty (e.) vector (ctr), the enzymatic dead construct K190M or the wild-type NSUN2 construct. NSUN2-specific sites are highlighted in red. d, e Heatmaps (upper panels) and methylation level (bottom panels) of VTRNA1.1, VTRN1.3, RPPH1, and HECTD1 in infected NSUN2-/- cells (d), human embryonic fibroblasts (H9) and HEK293 cells (e). Shown are five independent bisulfite conversion experiments. f Schematic illustration of NSUN2-dependent methylation (CH3) of VTRNA1.1 and the small regulatory non-coding fragments svRNA4. g Abundance of svRNA4 in the presence (+/−) and absence (−/−) of NSUN2. Methylation dead NSUN2-mutant constructs (C271A; C321A) fail to rescue svRNA4 levels in NSUN2−/− cells. Error bars indicate s.d. (n = 3 qRT-PCR reactions). ***p < 0.001 unpaired student’s t-test. h Log2 fold-change of NSUN2 in the indicated cells compared to NSUN2+/+control cells measured by ribosome profiling. Source data are provided as a Source Data file
