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 Concept Videos

Protein Complexes with Interchangeable Parts01:57

Protein Complexes with Interchangeable Parts

2.1K
Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
The SCF ubiquitin ligase is a protein complex of five individual proteins. This complex attaches ubiquitin to other target proteins to mark them for degradation. In order...
2.1K
Protein Networks02:26

Protein Networks

3.7K
An organism can have thousands of different proteins, and these proteins must cooperate to ensure the health of an organism. Proteins bind to other proteins and form complexes to carry out their functions. Many proteins interact with multiple other proteins creating a complex network of protein interactions.
These interactions can be represented through maps depicting protein-protein interaction networks, represented as nodes and edges. Nodes are circles that are representative of a protein,...
3.7K

You might also read

Related Articles

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

Sort by
Same author

KG-Microbe - Building Modular and Scalable Knowledge Graphs for Microbiome and Microbial Sciences.

GigaScienceĀ·2026
Same author

A quantitative metabolic signature of host response during SARS-CoV-2 infection and recovery.

iScienceĀ·2026
Same author

Esophageal epithelial cell-state transitions underlie the severity of pediatric eosinophilic esophagitis.

bioRxiv : the preprint server for biologyĀ·2026
Same author

OpenScientist: evaluating an open agentic AI co-scientist to accelerate biomedical discovery.

medRxiv : the preprint server for health sciencesĀ·2026
Same author

Systematic benchmarking demonstrates large language models have not reached the diagnostic accuracy of traditional rare-disease decision support tools.

European journal of human genetics : EJHGĀ·2026
Same author

Assessing the effectiveness of ontology-grounded AI term extraction using OntoGPT for environmental evidence synthesis.

Environmental evidenceĀ·2026

Related Experiment Video

Updated: May 2, 2026

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies
10:47

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

Published on: July 24, 2017

10.6K

Studying protein complexes by the yeast two-hybrid system.

Seesandra V Rajagopala1, Patricia Sikorski, J Harry Caufield

  • 1J Craig Venter Institute, Rockville, MD 20850, USA. rajgsv@gmail.com

Methods (San Diego, Calif.)
|July 31, 2012
PubMed
Summary

The yeast two-hybrid system reveals direct protein interactions within complexes. This method identified multiple interactions in Escherichia coli DNA polymerase III and other protein complexes, offering insights into complex topology.

More Related Videos

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

13.3K
Author Spotlight: Enhanced Isolation of Interaction-Null Mutants in Yeast
02:44

Author Spotlight: Enhanced Isolation of Interaction-Null Mutants in Yeast

Published on: December 29, 2023

1.1K

Related Experiment Videos

Last Updated: May 2, 2026

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies
10:47

A Modified Yeast-one Hybrid System for Heteromeric Protein Complex-DNA Interaction Studies

Published on: July 24, 2017

10.6K
A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions
14:23

A Yeast 2-Hybrid Screen in Batch to Compare Protein Interactions

Published on: June 6, 2018

13.3K
Author Spotlight: Enhanced Isolation of Interaction-Null Mutants in Yeast
02:44

Author Spotlight: Enhanced Isolation of Interaction-Null Mutants in Yeast

Published on: December 29, 2023

1.1K

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Structural Biology

Background:

  • Traditional methods like affinity purification and mass spectrometry often fail to elucidate the structure and topology of protein complexes.
  • Understanding direct protein-protein interactions is crucial for deciphering complex assembly and function.

Purpose of the Study:

  • To investigate the utility of the yeast two-hybrid (Y2H) system for analyzing direct interactions among proteins within complexes.
  • To assess the Y2H system's capability in mapping protein complex topology.

Main Methods:

  • Systematic pairwise interaction testing of proteins within the Escherichia coli DNA polymerase III complex.
  • Y2H analysis of an uncharacterized complex involving MntR and PerR.
  • Review of existing Y2H data for well-characterized complexes (VZV RNR, yeast proteasome, bacteriophage lambda) as gold standards.
  • Examination of Y2H analysis for the human spliceosome, a dynamic megacomplex.

Main Results:

  • Identified four direct interactions within the Escherichia coli DNA polymerase III complex.
  • Discovered seven direct interactions in the MntR/PerR-containing complex.
  • Validated Y2H approach using established protein complexes with known structures.
  • Demonstrated Y2H's potential for studying large, dynamic complexes like the human spliceosome.

Conclusions:

  • The yeast two-hybrid system is a powerful tool for identifying direct protein-protein interactions and inferring the topology of protein complexes.
  • Y2H analysis provides valuable insights complementary to traditional structural biology methods.
  • This approach is applicable to diverse biological systems, from simple bacterial complexes to complex eukaryotic machinery.