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

GCN4 binds with high affinity to DNA sequences containing a single consensus half-site.

J J Hollenbeck1, M G Oakley

  • 1Department of Chemistry, Indiana University, Bloomington 47405, USA.

Biochemistry
|June 1, 2000
PubMed
Summary

The bZip protein GCN4 can bind DNA as a dimer, even to sites with only one half-site. One monomer binds specifically, while the other makes important nonspecific contacts, with both basic regions becoming helical upon DNA binding.

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

Evaluation of the energetic contribution of interhelical Coulombic interactions for coiled coil helix orientation specificity.

Journal of molecular biology·2002
Same author

A GCN4 variant with a C-terminal basic region binds to DNA with wild-type affinity.

Biochemistry·2001
Same author

Selection of a high-affinity DNA pool for a bZip protein with an out-of-phase alignment of the basic region relative to the leucine zipper.

Bioorganic & medicinal chemistry·2001
Same author

The design of antiparallel coiled coils.

Current opinion in structural biology·2001
Same author

Design and characterization of a heterodimeric coiled coil that forms exclusively with an antiparallel relative helix orientation.

Journal of the American Chemical Society·2001
Same author

Developing and using preparatory information for women undergoing radiation therapy for cervical or uterine cancer.

Oncology nursing forum·2001

Area of Science:

  • Molecular Biology
  • Protein-DNA Interactions
  • Transcription Factor Regulation

Background:

  • bZip proteins are transcription factors with a bipartite DNA-binding motif.
  • They typically form dimeric complexes, with each monomer recognizing half of a DNA site.

Purpose of the Study:

  • Investigate the DNA binding mechanism of the bZip protein GCN4 to single half-sites.
  • Explore the structural basis of GCN4 binding to DNA, considering potential monomeric binding roles.

Main Methods:

  • Quantitative DNA binding assays
  • Affinity cleaving studies
  • Circular dichroism (CD) spectroscopy

Main Results:

  • GCN4 binds with high affinity to DNA sites with a single consensus half-site.

Related Experiment Videos

  • Binding occurs as a dimer, with one monomer specific and the other nonspecific DNA contact.
  • CD studies show basic regions of both monomers are helical upon DNA binding.

    • Conclusions:

    • GCN4 can bind DNA as a dimer to sites with single half-sites.
    • Nonspecific contacts by one monomer are crucial for binding affinity.
    • A protein folding transition, forming helical basic regions, is likely required for both specific and nonspecific DNA binding.