Related Experiment Videos
A high resolution protein interaction map of the yeast Mediator complex
Benjamin Guglielmi1, Nynke L van Berkum, Benjamin Klapholz
1Service de Biochimie et Génétique Moléculaire, Bâtiment 144, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France.
Nucleic Acids Research
|October 13, 2004
Summary
This study maps protein interactions within the yeast Mediator complex, revealing its conserved structure across species. The findings identify new subunit connections and confirm Mediator
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- The Mediator complex is a crucial transcriptional co-activator, mediating interactions between DNA-binding transcription factors and RNA polymerase II.
- In yeast (Saccharomyces cerevisiae), Mediator is a large complex comprising 24 subunits organized into head, middle, tail, and Cdk8 (Srb8-11) modules.
- Understanding Mediator subunit interactions is key to elucidating its regulatory mechanisms and structural organization.
Purpose of the Study:
- To investigate protein-protein contacts between budding yeast Mediator subunits.
- To construct a comprehensive interaction map of the yeast Mediator complex.
- To assess the evolutionary conservation of Mediator structural organization.
Main Methods:
- Utilized screening and pair-wise yeast two-hybrid assays to identify Mediator subunit interactions.
- Employed co-immunoprecipitation studies and gel filtration experiments for validation.
- Analyzed the protein interaction network within the Drosophila Mediator middle module.
Main Results:
- Generated an interaction map detailing numerous interaction domains between yeast Mediator subunits, many conserved evolutionarily.
- Discovered novel connections linking the Cdk8 (Srb8-11) and head modules, head and middle modules, and middle and tail modules.
- Confirmed Med31 (Soh1) association with the yeast Mediator, increasing its subunit count to 25.
- Demonstrated conserved structural organization of the Mediator complex from yeast to metazoans through Drosophila analysis.
Conclusions:
- The yeast Mediator complex comprises 25 subunits, including Med31 (Soh1).
- The established interaction map provides a framework for understanding Mediator structure-function relationships.
- Mediator complex organization is conserved across diverse eukaryotic species, highlighting its fundamental role in transcription regulation.