The Bacillus subtilis open reading frame ysgA encodes the SPOUT methyltransferase RlmP forming 2′-O-methylguanosine at position 2553 in the A-loop of 23S rRNA

DNAzyme-catalyzed cleavage of 23S rRNA is prevented by the 2′-O-methylation introduced by YsgA. Total RNA from B. subtilis wild-type (WT) or ΔysgA cells was incubated with a 34 base DNAzyme and analyzed by denaturing (8 M urea) 4% polyacrylamide gel electrophoresis, as described by Buchhaupt et al. (2007). The DNAzyme can cleave the 23S rRNA and generate 2579 and 349 nt fragments if G2553 is not 2′-O-methylated. The 2579 nt fragment is not separated from the full length 2928 nt 23S rRNA. The 349 nt fragment is indicated by an arrow.