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Control of SXT integration and excision.

Vincent Burrus1, Matthew K Waldor

  • 1Department of Microbiology and Medicine, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111, USA.

Journal of Bacteriology
|August 19, 2003
PubMed
Summary

The Vibrio cholerae SXT element integrates into the host chromosome via Int recombinase. A novel Xis protein facilitates SXT excision, a key step for transfer, though excised SXT doesn't guarantee transfer.

Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • The Vibrio cholerae SXT element is a mobile genetic element conferring antibiotic resistance.
  • SXT integrates into the host chromosome at a specific site (prfC) and can excise to a circular form.
  • Both integration and excision are mediated by the SXT-encoded Int recombinase.

Purpose of the Study:

  • To investigate the mechanisms of SXT integration and excision in Vibrio cholerae.
  • To identify factors regulating SXT mobility and transfer.
  • To characterize the role of Int and a novel protein, Xis, in SXT lifecycle.

Main Methods:

  • Site-specific recombination assays to study Int and Xis function.
  • Real-time quantitative PCR to measure SXT excision rates.

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  • Analysis of SXT integration at secondary attachment sites.

    • Main Results:

    • Int is necessary and sufficient for SXT integration, requiring activators SetC and SetD.
    • A novel SXT-encoded Xis protein acts as a recombination directionality factor, promoting excision and inhibiting integration.
    • SXT excision is required for transfer, but the presence of excised SXT does not correlate with donor activity.
    • SXT integration site influences excision and transfer efficiency, with pntB::SXT showing reduced transmissibility.

    Conclusions:

    • SXT integration and excision are complex processes regulated by specific proteins (Int, Xis, SetC, SetD).
    • Xis represents a new family of recombination directionality factors.
    • The integration site can impact SXT mobility, suggesting a link between genome structure and mobile element transfer.