Figure 1.
The amount of tRNA modifications in the mnm5(s2)U biosynthetic pathway is altered in the gidA mutant. (A) The pathway of mnm5(s2)U biosynthesis in E. coli. (B) Change in the levels of s2U, cmnm5U, and mnm5U in total tRNA isolated from wild-type, gidA mutant, and complemented strains. The wild type and gidA mutant carried the pBBR1MCS-4 plasmid as a vector control. The level of each modified ribonucleoside was quantified based on the MRM signal intensity and normalized by dividing the quantified amounts by the summed signals of adenosine, guanosine, cytidine, and uridine in the sample. The data represent the mean ± SD values of three biological replicates. The statistical analysis was performed using Graphpad Prism (GraphPad Software). Asterisks denote a significant difference in the one-way analysis of variance (ANOVA; * p≤ 0.05).