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Nucleotide sequences required for Tn3 transposition immunity.

J A Kans1, M J Casadaban

  • 1Department of Molecular Genetics and Cell Biology, University of Chicago, Illinois 60637.

Journal of Bacteriology
|April 1, 1989
PubMed
Summary
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Transposition immunity, where Tn3 transposons insert less frequently into DNA with existing copies, is mediated by terminal sequences. These sequences, specifically within the 38-base-pair inverted repeats, are crucial for this self-limiting genetic element behavior.

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • The Tn3 transposon exhibits transposition immunity, a phenomenon reducing its insertion frequency into DNA containing pre-existing Tn3 copies.
  • Understanding the molecular mechanisms underlying transposition immunity is crucial for controlling transposon activity and genome stability.

Purpose of the Study:

  • To identify and characterize the cis-acting DNA sequence responsible for mediating transposition immunity in the Tn3 transposon.
  • To elucidate the role of terminal sequences and palindromic regions within Tn3 in conferring immunity.

Main Methods:

  • Systematic deletion analysis of the Tn3 transposon's terminal regions to map the immunity-conferring sequence.
  • Plasmid-based insertion assays to quantify transposition frequency and assess the effects of deletions on immunity.

Related Experiment Videos

  • Sequence analysis to identify palindromic structures within the critical immunity region.

    • Main Results:

    • The cis-acting site for transposition immunity was localized to the terminal 38-base-pair sequence, which contains inversely repeated elements at the Tn3 ends.
    • Two palindromic sequences within this terminal region were identified as essential for immunity.
    • Deletions within this region resulted in partial or even negative immunity, while multiple copies of partially immune ends enhanced immunity.

    Conclusions:

    • The terminal 38-base-pair inverted repeat sequence of Tn3 is both necessary and sufficient for mediating transposition immunity.
    • Specific palindromic sequences within the terminal repeats play a critical role in the mechanism of transposition immunity.
    • No other regions of the Tn3 transposon are required for establishing this self-limiting insertion phenomenon.