Related Experiment Video

Updated: May 17, 2025

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
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DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

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RsmG methylation of 16S rRNA affects the function of ribosomal protein uS12

Trevor W Bell1, Rowan M Turner1, Amanda M Merryman1,2

  • 1Division of Biology and Biomedical Systems, School of Science and Engineering, University of Missouri-Kansas City, Rm 306 Spencer Hall5007 Rockhill Rd., Kansas City, MO, 64110, USA.

Archives of Microbiology
|May 16, 2025
PubMed

Insights

Inactivating the RsmG methyltransferase in Escherichia coli can lead to high-level streptomycin resistance, particularly when combined with specific mutations in ribosomal protein uS12. This finding reveals a new pathway for bacterial resistance to streptomycin.

Area of Science:

  • Bacteriology
  • Molecular Biology
  • Genetics

Background:

  • The RsmG methyltransferase is responsible for modifying G527 in bacterial 16S ribosomal RNA (rRNA).
  • Inactivation of RsmG typically confers low-level streptomycin resistance.
  • High-level streptomycin resistance usually involves mutations in ribosomal protein uS12 or 16S rRNA.

Purpose of the Study:

  • To investigate whether rsmG inactivation affects the streptomycin resistance phenotypes of Escherichia coli uS12 mutants.
  • To identify novel genetic combinations conferring high-level streptomycin resistance.

Main Methods:

  • Generation of random uS12 mutants in Escherichia coli.
  • Inactivation of the rsmG gene.
  • Assessment of streptomycin resistance (Minimum Inhibitory Concentration - MIC).
  • Genetic selections to identify streptomycin dependence.

Main Results:

  • While several uS12 mutants showed moderate streptomycin resistance (MIC 10-40 µg/ml) upon rsmG inactivation, one mutant (uS12 R85H) exhibited very high resistance (MIC >1,024 µg/ml).
  • Combined rsmG null mutations and specific uS12 alterations resulted in streptomycin dependence or pseudo-dependence.
  • Growth of some uS12 mutants on high streptomycin concentrations was conditional on rsmG inactivation.

Conclusions:

  • Loss of m7G527 methylation due to rsmG inactivation significantly impacts streptomycin resistance phenotypes in uS12 mutants.
  • This study identifies a novel mechanism for achieving high-level streptomycin resistance in bacteria through combined genetic alterations.
Keywords:
Ribosomal RNA methylaseRibosomal protein uS12RsmGStreptomycin resistance

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