TY - JOUR
T1 - Subunit architecture and functional modular rearrangements of the transcriptional mediator complex
AU - Tsai, Kuang Lei
AU - Tomomori-Sato, Chieri
AU - Sato, Shigeo
AU - Conaway, Ronald C.
AU - Conaway, Joan W.
AU - Asturias, Francisco J.
N1 - Funding Information:
This work was supported by US National Institutes of Health grants R01 GM67167 (F.J.A.) and R01 GM41628 (R.C.C. and J.W.C.) and by a grant to the Stowers Institute from the Helen Nelson Medical Research Fund at the Greater Kansas City Community Foundation. We thank Y. Takagi for providing the plasmid for MBP tagging in yeast, G.C. Lander for critical reading of manuscript, and Patrick Cramer for providing a Med19Δ yeast strain. We thank T. Otomo for providing the plasmid for HA-tagging in yeast. EM data collection was carried out at the National Resource for Automated Macromolecular Microscopy (NRAMM).
PY - 2014/6/5
Y1 - 2014/6/5
N2 - The multisubunit Mediator, comprising ∼30 distinct proteins, plays an essential role in gene expression regulation by acting as a bridge between DNA-binding transcription factors and the RNA polymerase II (RNAPII) transcription machinery. Efforts to uncover the Mediator mechanism have been hindered by a poor understanding of its structure, subunit organization, and conformational rearrangements. By overcoming biochemical and image analysis hurdles, we obtained accurate EM structures of yeast and human Mediators. Subunit localization experiments, docking of partial X-ray structures, and biochemical analyses resulted in comprehensive mapping of yeast Mediator subunits and a complete reinterpretation of our previous Mediator organization model. Large-scale Mediator rearrangements depend on changes at the interfaces between previously described Mediator modules, which appear to be facilitated by factors conducive to transcription initiation. Conservation across eukaryotes of Mediator structure, subunit organization, and RNA polymerase II interaction suggest conservation of fundamental aspects of the Mediator mechanism.
AB - The multisubunit Mediator, comprising ∼30 distinct proteins, plays an essential role in gene expression regulation by acting as a bridge between DNA-binding transcription factors and the RNA polymerase II (RNAPII) transcription machinery. Efforts to uncover the Mediator mechanism have been hindered by a poor understanding of its structure, subunit organization, and conformational rearrangements. By overcoming biochemical and image analysis hurdles, we obtained accurate EM structures of yeast and human Mediators. Subunit localization experiments, docking of partial X-ray structures, and biochemical analyses resulted in comprehensive mapping of yeast Mediator subunits and a complete reinterpretation of our previous Mediator organization model. Large-scale Mediator rearrangements depend on changes at the interfaces between previously described Mediator modules, which appear to be facilitated by factors conducive to transcription initiation. Conservation across eukaryotes of Mediator structure, subunit organization, and RNA polymerase II interaction suggest conservation of fundamental aspects of the Mediator mechanism.
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U2 - 10.1016/j.cell.2014.05.015
DO - 10.1016/j.cell.2014.05.015
M3 - Article
C2 - 24882805
AN - SCOPUS:84902186552
SN - 0092-8674
VL - 157
SP - 1430
EP - 1444
JO - Cell
JF - Cell
IS - 6
ER -