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

J. E. Peters, N. L. Craig

Research output: Contribution to journalArticlepeer-review

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.

Original languageEnglish (US)
Pages (from-to)737-747
Number of pages11
JournalGenes and Development
Volume15
Issue number6
DOIs
StatePublished - Mar 15 2001

Keywords

  • DNA replication
  • Target selection
  • Transposition

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

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