Figure 3.

CMAL is involved in the N₄-methylation of C₁₃₅₂ in the chloroplast 16S rRNA. (A) Alignment of 16S rDNA sequences from different species. The conserved nucleotides are enclosed in red lines. Blue asterisks indicate the C₁₃₅₂ site of the Arabidopsis 16S rDNA. (B) Methylation analysis of the C₁₃₅₂ site of the 16S rRNA. Total RNA from the WT and cmal was treated with sodium bisulfite and then subjected to RT-PCR with 16S-rRNA-specific oligonucleotides. The RT-PCR products were subcloned into T-vectors for sequencing. The PCR products generated using chloroplast genomic DNAs treated with sodium bisulfite were used as controls. The sequences were assayed using the Chrosome software. Blue asterisks indicate the C₁₃₅₂ site.