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Journal of Bacteriology, August 2007, p. 6068-6073, Vol. 189, No. 16
0021-9193/07/$08.00+0     doi:10.1128/JB.00558-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of the RsmG Methyltransferase Target as 16S rRNA Nucleotide G527 and Characterization of Bacillus subtilis rsmG Mutants{triangledown}

Kenji Nishimura,1,2 Shanna K. Johansen,3 Takashi Inaoka,1 Takeshi Hosaka,1 Shinji Tokuyama,2 Yasutaka Tahara,2 Susumu Okamoto,1 Fujio Kawamura,4 Stephen Douthwaite,3 and Kozo Ochi1*

National Food Research Institute, Tsukuba, Ibaraki 305-8642, Japan,1 Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University, Shizuoka 422-8529, Japan,2 Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense M, Denmark,3 Laboratory of Molecular Genetics and Research Information Center for Extremophiles, College of Science, Rikkyo University, Tokyo 171-8501, Japan4

Received 12 April 2007/ Accepted 5 June 2007

The methyltransferase RsmG methylates the N7 position of nucleotide G535 in 16S rRNA of Bacillus subtilis (corresponding to G527 in Escherichia coli). Disruption of rsmG resulted in low-level resistance to streptomycin. A growth competition assay revealed that there are no differences in fitness between the rsmG mutant and parent strains under the various culture conditions examined. B. subtilis rsmG mutants emerged spontaneously at a relatively high frequency, 10–6. Importantly, in the rsmG mutant background, high-level-streptomycin-resistant rpsL (encoding ribosomal protein S12) mutants emerged at a frequency 200 times greater than that seen for the wild-type strain. This elevated frequency in the emergence of high-level streptomycin resistance was facilitated by a mutation pattern in rpsL more varied than that obtained by selection of the wild-type strain.

* Corresponding author. Mailing address: National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki, 305-8642, Japan. Phone: 81-29-838-8125. Fax: 81-29-838-7996. E-mail: kochi{at}affrc.go.jp

{triangledown} Published ahead of print on 15 June 2007.

Journal of Bacteriology, August 2007, p. 6068-6073, Vol. 189, No. 16
0021-9193/07/$08.00+0     doi:10.1128/JB.00558-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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