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Conformational alterations in the ermC transcript in vivo during induction.

M Mayford1, B Weisblum

  • 1Department of Molecular Biology, University of Wisconsin, Madison 53706.

The EMBO Journal
|December 20, 1989
PubMed
Summary

The ermC gene

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Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • The ermC gene confers inducible antibiotic resistance in Staphylococcus aureus.
  • Its expression is regulated by mRNA secondary structure rearrangements.
  • This mechanism involves ribosome stalling during translation of a leader peptide.

Purpose of the Study:

  • To investigate the in vivo secondary structural changes of the ermC transcript during antibiotic induction.
  • To provide physical evidence for the proposed regulatory model of ermC expression.
  • To confirm the role of specific leader peptide codons in ermC induction.

Main Methods:

  • The ermC gene was expressed in Bacillus subtilis.
  • Dimethyl sulfate was used to probe mRNA structure in vivo.
  • Primer extension with reverse transcriptase identified modified bases.
  • Analysis focused on changes in mRNA secondary structure and ribosome protection sites.

Main Results:

  • Physical evidence confirmed secondary structure rearrangements in the ermC transcript upon induction.
  • Stalled ribosomes were shown to protect specific codons (9 and 10) of the leader peptide from chemical modification.
  • These findings align with previous genetic data on critical codons for induction.

Conclusions:

  • The study provides in vivo physical evidence supporting the mRNA secondary structure-based regulatory model for ermC.
  • Ribosome stalling at specific leader peptide codons is a key event in ermC induction.
  • This mechanism highlights a sophisticated control of antibiotic resistance gene expression.

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