Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Target DNA structure plays a critical role in Tn7 transposition.

P N Kuduvalli1, J E Rao, N L Craig

  • 1Howard Hughes Medical Institute, Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, 725 N. Wolfe Street, Baltimore, MD 21205, USA.

The EMBO Journal
|February 17, 2001
PubMed
Summary

The bacterial transposon Tn7 uses proteins TnsD and TnsC to bind attTn7 DNA. DNA distortion by TnsD signals TnsC binding, enabling site-specific Tn7 insertion.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Tn7: smarter than we thought.

Nature reviews. Molecular cell biology·2001
Same author

Selective recognition of pyrimidine motif triplexes by a protein encoded by the bacterial transposon Tn7.

Journal of molecular biology·2001
Same author

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

Genes & development·2001
Same author

Analysis of gain-of-function mutants of an ATP-dependent regulator of Tn7 transposition.

Journal of molecular biology·2001
Same author

Tn7 transposes proximal to DNA double-strand breaks and into regions where chromosomal DNA replication terminates.

Molecular cell·2000
Same author

Unexpected structural diversity in DNA recombination: the restriction endonuclease connection.

Molecular cell·2000

Area of Science:

  • Molecular Biology
  • Genetics
  • Microbiology

Background:

  • The bacterial transposon Tn7 mediates site-specific DNA integration using four proteins: TnsA, TnsB, TnsC, and TnsD.
  • Target site selection is primarily mediated by the TnsD protein binding to the attTn7 DNA sequence.

Purpose of the Study:

  • To elucidate the molecular mechanisms by which TnsD and TnsC interact with the attTn7 target site.
  • To understand how DNA distortion influences the recruitment and activation of the Tn7 transposition machinery.

Main Methods:

  • DNA-protein binding assays to determine interaction sites and grooves.
  • Analysis of DNA structural changes induced by protein binding.
  • Comparison of TnsD-attTn7 interactions with triplex DNA-mediated recruitment.

Related Experiment Videos

Main Results:

  • TnsD binding to attTn7 induces significant DNA distortion at the 5' end of the binding site.
  • TnsC interacts with the distorted DNA region and binds to the minor groove, while TnsD binds the major groove.
  • The TnsC-TnsD-attTn7 complex footprint encompasses the insertion site, with TnsC acting as a platform for transposase activation.

Conclusions:

  • DNA distortion at attTn7, induced by TnsD, acts as a crucial signal for TnsC recruitment.
  • The distinct DNA groove binding preferences of TnsD and TnsC are essential for precise target site recognition and transposition.
  • TnsC plays a central role in assembling and activating the Tn7 transposase complex at the specific attTn7 site.