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TitleA Pliable Mediator Acts as a Functional Rather Than an Architectural Bridge between Promoters and Enhancers.
Journal, issue, pagesCell, Vol. 178, Issue 5, Page 1145-1158.e20, Year 2019
Publish dateAug 22, 2019
AuthorsLaila El Khattabi / Haiyan Zhao / Jens Kalchschmidt / Natalie Young / Seolkyoung Jung / Peter Van Blerkom / Philippe Kieffer-Kwon / Kyong-Rim Kieffer-Kwon / Solji Park / Xiang Wang / Jordan Krebs / Subhash Tripathi / Noboru Sakabe / Débora R Sobreira / Su-Chen Huang / Suhas S P Rao / Nathanael Pruett / Daniel Chauss / Erica Sadler / Andrea Lopez / Marcelo A Nóbrega / Erez Lieberman Aiden / Francisco J Asturias / Rafael Casellas /
PubMed AbstractWhile Mediator plays a key role in eukaryotic transcription, little is known about its mechanism of action. This study combines CRISPR-Cas9 genetic screens, degron assays, Hi-C, and cryoelectron ...While Mediator plays a key role in eukaryotic transcription, little is known about its mechanism of action. This study combines CRISPR-Cas9 genetic screens, degron assays, Hi-C, and cryoelectron microscopy (cryo-EM) to dissect the function and structure of mammalian Mediator (mMED). Deletion analyses in B, T, and embryonic stem cells (ESC) identified a core of essential subunits required for Pol II recruitment genome-wide. Conversely, loss of non-essential subunits mostly affects promoters linked to multiple enhancers. Contrary to current models, however, mMED and Pol II are dispensable to physically tether regulatory DNA, a topological activity requiring architectural proteins. Cryo-EM analysis revealed a conserved core, with non-essential subunits increasing structural complexity of the tail module, a primary transcription factor target. Changes in tail structure markedly increase Pol II and kinase module interactions. We propose that Mediator's structural pliability enables it to integrate and transmit regulatory signals and act as a functional, rather than an architectural bridge, between promoters and enhancers.
External linksCell / PubMed:31402173 / PubMed Central
MethodsEM (single particle)
Resolution4.02 - 8.28 Å
Structure data

EMDB-20391:
CryoEM structure of mouse mediator subunit one deletion
Method: EM (single particle) / Resolution: 8.28 Å

EMDB-20392:
CryoEM structure of wild type mouse mediator with MED19-FLAG
Method: EM (single particle) / Resolution: 5.88 Å

EMDB-20393:
CryoEM structure of wild type mediator with MED25-FLAG
Method: EM (single particle) / Resolution: 5.49 Å

21514

6w1s

EMDB-21514: map of the mammalian Mediator complex
PDB-6w1s: Atomic model of the mammalian Mediator complex
Method: EM (single particle) / Resolution: 4.02 Å

Source
  • mus musculus (house mouse)
KeywordsGENE REGULATION / mouse mediator / complex

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