TY - JOUR
T1 - A high resolution protein interaction map of the yeast Mediator complex
AU - Guglielmi, Benjamin
AU - van Berkum, Nynke L.
AU - Klapholz, Benjamin
AU - Bijma, Theo
AU - Boube, Muriel
AU - Boschiero, Claire
AU - Bourbon, Henri Marc
AU - Holstege, Frank C.P.
AU - Werner, Michel
N1 - Funding Information:
We thank M. Fromont-Racine for providing us with the FRYL2 library, B. Termenon and D. Tourbiez for technical help, A. Sentenac and P. Thuriaux for careful reading of the manuscript. H.-M.B acknowledges David Cribbs for his constant support. B.G. was supported by grants from the Ministère de la Recherche et de la Technologie and from the Association pour la Recherche sur le Cancer (ARC). This work was supported from a grant (to H.-M.B.) from the ARC. N.B., F.H. and T.B. are supported by grants from the Netherlands Organization for Scientific Research (NWO).
PY - 2004
Y1 - 2004
N2 - Mediator is a large, modular protein complex remotely conserved from yeast to man that conveys regulatory signals from DNA-binding transcription factors to RNA polymerase II. In Saccharomyces cerevisiae, Mediator is thought to be composed of 24 subunits organized in four sub-complexes, termed the head, middle, tail and Cdk8 (Srb8-11) modules. In this work, we have used screening and pair-wise two-hybrid approaches to investigate protein-protein contacts between budding yeast Mediator subunits. The derived interaction map includes the delineation of numerous interaction domains between Mediator subunits, frequently corresponding to segments that have been conserved in evolution, as well as novel connections between the Cdk8 (Srb8-11) and head modules, the head and middle modules, and the middle and tail modules. The two-hybrid analysis, together with co-immunoprecipitation studies and gel filtration experiments revealed that Med31 (Soh1) is associated with the yeast Mediator that therefore comprises 25 subunits. Finally, analysis of the protein interaction network within the Drosophila Mediator middle module indicated that the structural organization of the Mediator complex is conserved from yeast to metazoans. The resulting interaction map provides a framework for delineating Mediator structure-function and investigating how Mediator function is regulated.
AB - Mediator is a large, modular protein complex remotely conserved from yeast to man that conveys regulatory signals from DNA-binding transcription factors to RNA polymerase II. In Saccharomyces cerevisiae, Mediator is thought to be composed of 24 subunits organized in four sub-complexes, termed the head, middle, tail and Cdk8 (Srb8-11) modules. In this work, we have used screening and pair-wise two-hybrid approaches to investigate protein-protein contacts between budding yeast Mediator subunits. The derived interaction map includes the delineation of numerous interaction domains between Mediator subunits, frequently corresponding to segments that have been conserved in evolution, as well as novel connections between the Cdk8 (Srb8-11) and head modules, the head and middle modules, and the middle and tail modules. The two-hybrid analysis, together with co-immunoprecipitation studies and gel filtration experiments revealed that Med31 (Soh1) is associated with the yeast Mediator that therefore comprises 25 subunits. Finally, analysis of the protein interaction network within the Drosophila Mediator middle module indicated that the structural organization of the Mediator complex is conserved from yeast to metazoans. The resulting interaction map provides a framework for delineating Mediator structure-function and investigating how Mediator function is regulated.
UR - http://www.scopus.com/inward/record.url?scp=6044249068&partnerID=8YFLogxK
U2 - 10.1093/nar/gkh878
DO - 10.1093/nar/gkh878
M3 - Article
C2 - 15477388
AN - SCOPUS:6044249068
SN - 0305-1048
VL - 32
SP - 5379
EP - 5391
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 18
ER -