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Related Experiment Videos

Tipping the balance between replicative and simple transposition.

N P Tavakoli1, K M Derbyshire

  • 1Division of Infectious Disease, Wadsworth Center, New York State Department of Health, and School of Public Health, State University of New York at Albany, Albany, NY 12201-2002, USA.

The EMBO Journal
|June 2, 2001
PubMed
Summary
This summary is machine-generated.

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Bacterial insertion sequence IS903 transposition can be replicative or non-replicative. Flanking DNA and transposase mutations significantly alter transposition outcomes, favoring co-integrate formation.

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • The bacterial insertion sequence IS903 exhibits dual transposition modes: replicative and non-replicative.
  • Replicative transposition, forming co-integrates, is rare (<0.1% of simple insertions).

Purpose of the Study:

  • To identify critical steps determining IS903 transposition outcome.
  • To isolate mutants enhancing replicative transposition frequency.

Main Methods:

  • Isolation and characterization of IS903 mutants affecting transposition.
  • Analysis of flanking nucleotide effects on transposition frequency and co-integrate formation.
  • Identification of transposase mutations impacting replicative transposition.

Main Results:

Related Experiment Videos

  • The 3'-flanking nucleotide significantly influences transposition frequency and co-integrate formation; a 3'-A increases co-integrates 500-fold over 3'-C.
  • Five transposase mutants were identified that increase replicative transposition.
  • These mutations are located near catalytic residues, suggesting active site involvement.

Conclusions:

  • Transposase mutations can specifically alter transposition product outcome.
  • A delayed 5'-flanking DNA cleavage enhances co-integrate formation by stabilizing the 3'-nicked intermediate.