Target DNA structure plays a critical role in Tn7 transposition

P. N. Kuduvalli, J. E. Rao, Nancy L Craig

Research output: Contribution to journalArticle

Abstract

The bacterial transposon Tn7 utilizes four Tn7-encoded proteins, TnsA, TnsB, TnsC and TnsD, to make insertions at a specific site termed attTn7. This target is selected by the binding of TnsD to attTn7 in a sequence-specific manner, followed by the binding of TnsC and activation of the transposase. We show that TnsD binding to attTn7 induces a distortion at the 5′ end of the binding site and TnsC contacts the region of attTn7 distorted by TnsD. Previous work has shown that a target site containing triplex DNA, instead of TnsD-attTn7, can recruit TnsABC and effect site-specific insertion of Tn7. We propose that the DNA distortion imposed by TnsD on attTn7, like the altered DNA structure via triplex formation, serves as a signal to recruit TnsC. We also show that TnsD primarily contacts the major groove of DNA, whereas TnsC is a minor groove binding protein. The footprint of the TnsC-TnsD-attTn7 nucleoprotein complex includes and extends beyond the Tn7 insertion site, where TnsC forms a platform to receive and activate the transposase to carry out recombination.

Original languageEnglish (US)
Pages (from-to)924-932
Number of pages9
JournalThe EMBO journal
Volume20
Issue number4
DOIs
StatePublished - Feb 15 2001

Fingerprint

Transposases
DNA
Nucleoproteins
Genetic Recombination
Carrier Proteins
Chemical activation
Binding Sites
Proteins
triplex DNA

Keywords

  • DNA distortion
  • Footprinting
  • Protein-DNA
  • Recombination
  • Triplex

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

Target DNA structure plays a critical role in Tn7 transposition. / Kuduvalli, P. N.; Rao, J. E.; Craig, Nancy L.

In: The EMBO journal, Vol. 20, No. 4, 15.02.2001, p. 924-932.

Research output: Contribution to journalArticle

Kuduvalli, P. N. ; Rao, J. E. ; Craig, Nancy L. / Target DNA structure plays a critical role in Tn7 transposition. In: The EMBO journal. 2001 ; Vol. 20, No. 4. pp. 924-932.
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