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Mutational analysis of basic residues in the N-terminus of the rRNA:m6A methyltransferase ErmC′

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Abstract

Erm methyltransferases mediate the resistance to the macrolide-lincosamide-streptogramin B antibioticsvia dimethylation of a specific adenine residue in 23S rRNA. The role of positively charged N-terminal residues of the ErmC′ methyltransferase in RNA binding and/or catalysis was determined. Mutational analysis of amino acids K4 and K7 was performed and the mutants were characterized inin vivo andin vitro experiments. The K4 and K7 residues were suggested not to be essential for the enzyme activity but to provide a considerable support for the catalytic step of the reaction, probably by maintaining the optimum conformation of the transition state through interactions with the phosphate backbone of RNA.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Faculty of Pharmacy and Biochemistry, University of Zagreb, 10000, Zagreb, Croatia

    G. Maravić & M. Flögel

  2. Bioinformatics Laboratory, International Institute of Molecular and Cell Biology, 02-109, Warsaw, Poland

    J. M. Bujnicki

Authors
  1. G. Maravić
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  2. J. M. Bujnicki
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  3. M. Flögel
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Correspondence to G. Maravić.

Additional information

This work was supported by theMinistry of Science of Republic Croatia (no. 006320 and 006611). The second author is theEuropean Molecular Biology Organization andHoward Hughes Medical Institute young investigator and theFoundation for Polish Science fellow.

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Maravić, G., Bujnicki, J.M. & Flögel, M. Mutational analysis of basic residues in the N-terminus of the rRNA:m6A methyltransferase ErmC′. Folia Microbiol 49, 3–7 (2004). https://doi.org/10.1007/BF02931637

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  • DOI: https://doi.org/10.1007/BF02931637

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