Faculty, Staff and Student Publications

Language

English

Publication Date

10-17-2024

Journal

Molecular Cell

DOI

10.1016/j.molcel.2024.09.001

PMID

39321804

PMCID

PMC11832219

PubMedCentral® Posted Date

10-17-2025

PubMedCentral® Full Text Version

Author MSS

Abstract

The eukaryotic transcriptional Mediator comprises a large core (cMED) and a dissociable CDK8 kinase module (CKM). cMED recruits RNA polymerase II (RNA Pol II) and promotes pre-initiation complex formation in a manner repressed by the CKM through mechanisms presently unknown. Herein, we report cryoelectron microscopy structures of the complete human Mediator and its CKM. The CKM binds to multiple regions on cMED through both MED12 and MED13, including a large intrinsically disordered region (IDR) in the latter. MED12 and MED13 together anchor the CKM to the cMED hook, positioning CDK8 downstream and proximal to the transcription start site. Notably, the MED13 IDR obstructs the recruitment of RNA Pol II/MED26 onto cMED by direct occlusion of their respective binding sites, leading to functional repression of cMED-dependent transcription. Combined with biochemical and functional analyses, these structures provide a conserved mechanistic framework to explain the basis for CKM-mediated repression of cMED function.

Keywords

Humans, Mediator Complex, Cyclin-Dependent Kinase 8, Cryoelectron Microscopy, RNA Polymerase II, Binding Sites, Protein Binding, Transcription, Genetic, Models, Molecular, Structure-Activity Relationship, Intrinsically Disordered Proteins

Published Open-Access

yes

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