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An IS26 variant with enhanced activity.

Carol H Pong1, Christopher J Harmer1, Sandro F Ataide1

  • 1School of Life and Environmental Sciences, Molecular Bioscience Building G08, Cnr Maze Crescent and Butlin Avenue, The University of Sydney, NSW 2006, Australia.

FEMS Microbiology Letters
|February 13, 2019
PubMed
Summary
This summary is machine-generated.

The common variant IS26* transposase (Tnp26-G184N) significantly enhances the mobility of antibiotic resistance genes in bacteria. This G184N substitution boosts transposition activity, aiding the spread of antimicrobial resistance.

Keywords:
antibiotic resistancecointegrateinsertion sequencetransposon

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • The insertion sequence IS26 is crucial for the spread of antibiotic resistance genes in Gram-negative bacteria.
  • Despite IS26's abundance, few sequence variations have been documented.
  • The common variant IS26* encodes a Tnp26 transposase with a G184N substitution in its catalytic domain.

Purpose of the Study:

  • To investigate the impact of the G184N substitution in Tnp26 on IS26 transposition activity.
  • To understand how this specific amino acid change affects the catalytic domain and DNA binding.

Main Methods:

  • Computational modeling to predict the structural effects of the G184N substitution.
  • In vitro transposition assays using wild-type IS26 and the G184N mutant.

Main Results:

  • The G184N substitution increased cointegrate formation 5-fold in standard reactions.
  • Targeted transposition assays showed a 10-fold increase in cointegration frequency with the G184N mutant.
  • Computational modeling suggested the substitution lengthens a helix, potentially stabilizing the catalytic triad.

Conclusions:

  • The G184N substitution significantly enhances Tnp26 activity, increasing transposition efficiency.
  • Structural changes, including helix stabilization and improved binding to terminal repeats, likely underlie the enhanced activity.
  • This finding provides insight into the mechanisms driving the dissemination of antibiotic resistance genes.