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Tn7 recognizes transposition target structures associated with DNA replication using the DNA-binding protein TnsE.

J E Peters·N L Craig
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Abstract

We report that the bacterial transposon Tn7 selects targets by recognizing features associated with DNA replication using the transposon-encoded DNA-binding protein TnsE. We show that Tn7 transposition directed by TnsE occurs in one orientation with respect to chromosomal DNA replication, indicating that a structure or complex involved in DNA replication is likely to be a critical determinant of TnsE insertion. We find that mutant TnsE proteins that allow higher levels of transposition also bind DNA better than the wild-type protein. The increased binding affinity displayed by the TnsE high-activity mutants indicates that DNA binding is relevant to transposition activity and suggests that TnsE interacts directly with target DNAs. In vitro, TnsE interacts preferentially with certain DNA structures, indicating a mechanism for the TnsE-mediated orientation and insertion preference. The pattern of TnsE-mediated insertion events around the Escherichia coli chromosome provides insight into how DNA replication forks proceed in vivo.

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AllelesAmino Acid SequenceBlotting, WesternDNADNA ReplicationDNA Transposable ElementsDNA-Binding ProteinsEscherichia coliEscherichia coli ProteinsModels, GeneticMolecular Sequence DataMutationPlasmidsPolymerase Chain ReactionProtein BindingSequence Homology, Amino AcidJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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J E Peters, N L Craig. 2001-03-15. Tn7 recognizes transposition target structures associated with DNA replication using the DNA-binding protein TnsE.. https://doi.org/10.1101/gad.870201

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