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Related Experiment Videos

Multiple DNA processing reactions underlie Tn7 transposition

P A Gary1, M C Biery, R J Bainton

  • 1Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, John Hopkins Universtiy School of Medicine, Baltimore, MD 21205, USA.

Journal of Molecular Biology
|March 29, 1996
PubMed
Summary
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The bacterial transposon Tn7 uses a cut and paste mechanism for DNA translocation. Its transposition involves distinct DNA processing steps, including cleavage and strand transfer at the 3' ends and cleavage at the 5' ends.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The bacterial transposon Tn7 facilitates its movement between DNA sites using a cut-and-paste mechanism.
  • This process involves creating double-strand breaks at transposon ends and joining the transposon to target DNA.

Purpose of the Study:

  • To dissect the chemical steps involved in Tn7 transposition.
  • To investigate the roles of DNA strand processing activities at transposon ends.

Main Methods:

  • Utilized mutant transposons with alterations at and near their termini.
  • Analyzed the effects of these mutations on DNA cleavage and joining reactions.

Main Results:

  • Identified two distinct DNA strand processing activities responsible for initiating double-strand breaks at transposon ends.

Related Experiment Videos

  • Demonstrated that changes at transposon tips can block both cleavage and joining steps.
  • Showed that target joining occurs via two separable reactions involving individual 3' transposon ends joining separate target DNA strands.

    • Conclusions:

    • Tn7 transposition involves multiple distinct DNA processing reactions.
    • These reactions include strand cleavage and strand transfer at the 3' ends, and separate strand cleavage at the 5' ends.
    • The target joining step can be dissected into separable reactions, highlighting the complexity of transposon integration.