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Related Experiment Videos

Switching from cut-and-paste to replicative Tn7 transposition

E W May1, N L Craig

  • 1Howard Hughes Medical Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21205 USA.

Science (New York, N.Y.)
|April 19, 1996
PubMed
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A single mutation in the bacterial transposon Tn7 alters its transposition mechanism from cut-and-paste to replicative. This switch results from blocking 5' end processing, revealing Tn7's adaptable DNA repair and replication strategies.

Area of Science:

  • * Molecular Biology
  • * Genetics
  • * Microbiology

Background:

  • * The bacterial transposon Tn7 typically utilizes a cut-and-paste mechanism for transposition.
  • * This process involves excising the transposon from a donor DNA site and inserting it into a target site, forming a simple insertion.

Purpose of the Study:

  • * To investigate the mechanism by which Tn7 transposition can switch from a cut-and-paste to a replicative mode.
  • * To identify the specific molecular events that lead to this mechanistic switch.

Main Methods:

  • * Investigated Tn7 transposition in vitro and in vivo.
  • * Introduced a specific amino acid alteration in a Tn7-encoded protein essential for transposition.
  • * Analyzed the processing of transposon ends and the resulting DNA products.

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Main Results:

  • * A single amino acid mutation in a key Tn7 protein blocked 5' strand cleavage but not 3' strand processing.
  • * This selective blockade resulted in the formation of a Shapiro Intermediate, leading to replicative transposition.
  • * The study demonstrated the generation of plasmid fusions in vitro and cointegrate products in vivo.

Conclusions:

  • * The findings reveal that separating 5' and 3' end processing reactions dictates the transposition pathway of Tn7.
  • * Tn7 exhibits remarkable plasticity in utilizing host DNA repair and replication machinery for both cut-and-paste and replicative transposition.