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Related Experiment Videos

Trans-acting transposase mutant from Tn5.

A DeLong1, M Syvanen

  • 1Department of Biology, Yale University, New Haven, CT 06511.

Proceedings of the National Academy of Sciences of the United States of America
|July 15, 1991
PubMed
Summary
This summary is machine-generated.

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A novel mutant transposase (Tnp-449F) from the Tn5 transposon system can act in trans, unlike the wild-type enzyme. This discovery sheds light on the regulation of transposition and protein function.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Transposition of Tn5 and IS50R is regulated by transposase (Tnp) and inhibitor (Inh) proteins.
  • Understanding the cis-acting mechanism of Tnp and the role of inhibition is crucial.

Purpose of the Study:

  • To investigate the mechanism of cis-acting Tnp and the relevance of inhibition to cis action.
  • To screen for mutants with altered transposition regulation.

Main Methods:

  • Utilized a colony morphology assay sensitive to Inh for screening transposition mutants.
  • Isolated and characterized a dominant mutant (449F) in IS50R promoting trans-transposition.

Main Results:

  • Identified a Leu-Phe mutation at position 449 in Tnp (Tnp-449F).

Related Experiment Videos

  • Tnp-449F reduces cis-transposition but acts efficiently in trans.
  • Tnp-449F is sensitive to inhibition, and Inh-449F is a competent inhibitor.

    • Conclusions:

    • Wild-type Tnp is cis-acting, while the mutant Tnp-449F exhibits trans-acting transposase activity.
    • This finding provides new insights into the regulatory mechanisms of Tn5 transposition.