Sequence Analysis and Structure Prediction of 23S rRNA: m~1G Methyltransferases Reveals a Conserved Core Augmented with a Putative Zn-binding Domain in the N-Terminus and Family-Specific Elaborations in the C-Terminus

摘要

N1-methylation of G748 within 23S ribosomal RNA results in resistance to the macrolide tylosin in Streptomyces. In contrast, the Escherichia coli mutant lacking N1-methylation of G745 exhibits increased resistance to viomycin, in addition to severe defects of growth characteristics. Both methylated guanines are located in hairpin 35, in domain II of prokaryotic 23S rRNA. G748 and G745 are modified by related S-adenosylmethionine-dependent methyltransferases (MTases), TlrB and RrmA respectively. Earlier sequence comparisons allowed identification of the AdoMet-binding site, however the catalytic site and the target-recognition region of these enzyme could not be delineated unambiguously. In this work, we carried out sequence-to-structure threading of the rRNA:m~1G MTase family against the database of known structures to identify those "missing regions". Our analysis confirms the earlier prediction of the AdoMet-binding site, but suggests a different location of the putative catalytic center than was previously postulated. We predict that RrmA and TlrB possess two regions that may be responsible for specific interactions with their target nucleic acid sequences: a putative Zn-finger domain in the N-terminus and the variable domain close to the C-terminus, which indicates that 23S rRNA MTases from other nucleic acid MTases, despite sharing the common catalytic domain.