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- PDB-6w1s: Atomic model of the mammalian Mediator complex -

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Entry
Database: PDB / ID: 6w1s
TitleAtomic model of the mammalian Mediator complex
Components(Mediator of RNA polymerase II transcription subunit ...) x 25
KeywordsGENE REGULATION / mouse mediator / complex
Function / homology
Function and homology information


: / Generic Transcription Pathway / regulation of cell cycle => GO:0051726 / positive regulation of mediator complex assembly / positive regulation of T cell extravasation / negative regulation of smooth muscle cell differentiation / core mediator complex / mediator complex / nuclear retinoic acid receptor binding / positive regulation of chromatin binding ...: / Generic Transcription Pathway / regulation of cell cycle => GO:0051726 / positive regulation of mediator complex assembly / positive regulation of T cell extravasation / negative regulation of smooth muscle cell differentiation / core mediator complex / mediator complex / nuclear retinoic acid receptor binding / positive regulation of chromatin binding / limb development / nuclear thyroid hormone receptor binding / termination of RNA polymerase II transcription / cortical actin cytoskeleton / stem cell population maintenance / skeletal muscle cell differentiation / blastocyst development / transcription factor binding / positive regulation of transcription initiation by RNA polymerase II / ubiquitin ligase complex / nuclear retinoid X receptor binding / catalytic activity / negative regulation of fibroblast proliferation / histone acetyltransferase activity / nuclear receptor coactivator activity / transcription coregulator activity / transcription coactivator binding / ubiquitin protein ligase activity / actin binding / transcription regulator complex / transcription by RNA polymerase II / transcription coactivator activity / protein ubiquitination / nuclear body / cytoskeleton / RNA polymerase II cis-regulatory region sequence-specific DNA binding / protein-containing complex binding / positive regulation of gene expression / nucleolus / regulation of DNA-templated transcription / regulation of transcription by RNA polymerase II / positive regulation of DNA-templated transcription / negative regulation of transcription by RNA polymerase II / positive regulation of transcription by RNA polymerase II / DNA binding / nucleoplasm / membrane / nucleus / cytosol / cytoplasm
Similarity search - Function
Mediator complex, subunit Med25, NR box / : / : / Mediator complex, subunit Med25, PTOV domain / Mediator complex, subunit Med25, synapsin 1 / Mediator of RNA polymerase II transcription subunit 25, von Willebrand factor type A domain / Mediator complex subunit 25, PTOV domain superfamily / Mediator complex subunit 25 PTOV activation and synapsin 2 / Mediator complex subunit 25 synapsin 1 / Mediator complex subunit 25 von Willebrand factor type A ...Mediator complex, subunit Med25, NR box / : / : / Mediator complex, subunit Med25, PTOV domain / Mediator complex, subunit Med25, synapsin 1 / Mediator of RNA polymerase II transcription subunit 25, von Willebrand factor type A domain / Mediator complex subunit 25, PTOV domain superfamily / Mediator complex subunit 25 PTOV activation and synapsin 2 / Mediator complex subunit 25 synapsin 1 / Mediator complex subunit 25 von Willebrand factor type A / Mediator complex, subunit Med29, metazoa / Mediator complex, subunit Med24, / Mediator complex subunit 24 N-terminal / Mediator complex subunit 29 / Mediator of RNA polymerase II transcription subunit 15 / Mediator complex, subunit Med23 / ARC105/Med15 mediator subunit N-terminal KIX domain / Mediator complex subunit 23 / Mediator complex subunit 28 / Mediator complex, subunit Med6, metazoa/plant / Mediator complex, subunit Med19, metazoa / Mediator complex, subunit Med30, metazoa / Mediator complex, subunit Med28 / Mediator of RNA polymerase II transcription subunit 9, metazoan / Mediator of RNA pol II transcription subunit 19 / Mediator complex subunit 30 / Mediator complex, subunit Med27 / Mediator complex subunit 27 / Mediator complex, subunit Med16 / Mediator complex subunit 16, N-terminal / Mediator complex, subunit Med10 / Transcription factor subunit Med10 of Mediator complex / Mediator of RNA polymerase II transcription subunit 9 / RNA polymerase II transcription mediator complex subunit 9 / Mediator complex, subunit Med4 / Vitamin-D-receptor interacting Mediator subunit 4 / Mediator complex, subunit Med31 / Mediator complex, subunit Med21 / Mediator of RNA polymerase II, subunit Med31 superfamily / SOH1 / Subunit 21 of Mediator complex / Mediator complex, subunit Med7 / Soluble quinoprotein glucose/sorbosone dehydrogenase / Mediator complex, subunit Med7/Med21-like / MED7 protein / Mediator complex, subunit Med20 / Mediator complex, subunit Med14 / TATA-binding related factor (TRF) of subunit 20 of Mediator complex / Mediator complex subunit MED14 / Mediator complex, subunit Med6 / Mediator complex, subunit Med17 / Mediator complex, subunit Med6 superfamily / MED6 mediator sub complex component / Subunit 17 of Mediator complex / Mediator of RNA polymerase II transcription subunit 22 / Mediator complex, subunit Med8, fungi/metazoa / Mediator complex, subunit Med11 / Surfeit locus protein 5 subunit 22 of Mediator complex / Mediator of RNA polymerase II transcription complex subunit 8 / Mediator complex protein / Mediator complex, subunit Med18 / Med18 protein / : / Coactivator CBP, KIX domain superfamily / von Willebrand factor A-like domain superfamily / WD40 repeats / WD40 repeat / Trp-Asp (WD) repeats profile. / Trp-Asp (WD) repeats circular profile. / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
Similarity search - Domain/homology
Mediator of RNA polymerase II transcription subunit 14 / Mediator of RNA polymerase II transcription subunit 22 / Mediator of RNA polymerase II transcription subunit 16 / Mediator of RNA polymerase II transcription subunit 23 / Mediator of RNA polymerase II transcription subunit 19 / Mediator of RNA polymerase II transcription subunit 25 / Mediator of RNA polymerase II transcription subunit 17 / Mediator of RNA polymerase II transcription subunit 9 / Mediator of RNA polymerase II transcription subunit 28 / Mediator of RNA polymerase II transcription subunit 6 ...Mediator of RNA polymerase II transcription subunit 14 / Mediator of RNA polymerase II transcription subunit 22 / Mediator of RNA polymerase II transcription subunit 16 / Mediator of RNA polymerase II transcription subunit 23 / Mediator of RNA polymerase II transcription subunit 19 / Mediator of RNA polymerase II transcription subunit 25 / Mediator of RNA polymerase II transcription subunit 17 / Mediator of RNA polymerase II transcription subunit 9 / Mediator of RNA polymerase II transcription subunit 28 / Mediator of RNA polymerase II transcription subunit 6 / Mediator of RNA polymerase II transcription subunit 15 / Mediator of RNA polymerase II transcription subunit 24 / Mediator of RNA polymerase II transcription subunit 21 / Mediator of RNA polymerase II transcription subunit 4 / Mediator of RNA polymerase II transcription subunit 30 / Mediator of RNA polymerase II transcription subunit 10 / Mediator of RNA polymerase II transcription subunit 31 / Mediator of RNA polymerase II transcription subunit 18 / Mediator of RNA polymerase II transcription subunit 7 / Mediator of RNA polymerase II transcription subunit 8 / Mediator of RNA polymerase II transcription subunit 11 / Mediator of RNA polymerase II transcription subunit 27 / Mediator of RNA polymerase II transcription subunit 29 / Mediator of RNA polymerase II transcription subunit 20
Similarity search - Component
Biological speciesMus musculus (house mouse)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 4.02 Å
AuthorsZhao, H. / Young, N. / Asturias, F.
Funding support United States, 2items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)R01-67167 United States
National Institutes of Health/National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIH/NIAMS) United States
CitationJournal: Cell / Year: 2019
Title: A Pliable Mediator Acts as a Functional Rather Than an Architectural Bridge between Promoters and Enhancers.
Authors: Laila 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 ...Authors: Laila 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 /
Abstract: 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 ...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.
History
DepositionMar 4, 2020Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 10, 2021Provider: repository / Type: Initial release

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Structure visualization

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Assembly

Deposited unit
A: Mediator of RNA polymerase II transcription subunit 1
B: Mediator of RNA polymerase II transcription subunit 4
C: Mediator of RNA polymerase II transcription subunit 6
D: Mediator of RNA polymerase II transcription subunit 7
E: Mediator of RNA polymerase II transcription subunit 8
F: Mediator of RNA polymerase II transcription subunit 9
G: Mediator of RNA polymerase II transcription subunit 10
H: Mediator of RNA polymerase II transcription subunit 11
I: Mediator of RNA polymerase II transcription subunit 14
J: Mediator of RNA polymerase II transcription subunit 15
K: Mediator of RNA polymerase II transcription subunit 16
L: Mediator of RNA polymerase II transcription subunit 17
M: Mediator of RNA polymerase II transcription subunit 18
N: Mediator of RNA polymerase II transcription subunit 19
O: Mediator of RNA polymerase II transcription subunit 20
P: Mediator of RNA polymerase II transcription subunit 21
Q: Mediator of RNA polymerase II transcription subunit 22
R: Mediator of RNA polymerase II transcription subunit 23
S: Mediator of RNA polymerase II transcription subunit 24
T: Mediator of RNA polymerase II transcription subunit 25
V: Mediator of RNA polymerase II transcription subunit 27
W: Mediator of RNA polymerase II transcription subunit 28
X: Mediator of RNA polymerase II transcription subunit 29
Y: Mediator of RNA polymerase II transcription subunit 30
Z: Mediator of RNA polymerase II transcription subunit 31


Theoretical massNumber of molelcules
Total (without water)975,06725
Polymers975,06725
Non-polymers00
Water0
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_5551

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Components

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Mediator of RNA polymerase II transcription subunit ... , 25 types, 25 molecules ABCDEFGHIJKLMNOPQRSTVWXYZ

#1: Protein Mediator of RNA polymerase II transcription subunit 1


Mass: 32527.998 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse)
#2: Protein Mediator of RNA polymerase II transcription subunit 4 / Mediator complex subunit 4


Mass: 11647.339 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CQA5
#3: Protein Mediator of RNA polymerase II transcription subunit 6 / Mediator complex subunit 6


Mass: 28473.553 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q921D4
#4: Protein Mediator of RNA polymerase II transcription subunit 7 / Cofactor required for Sp1 transcriptional activation subunit 9 / CRSP complex subunit 9 / Mediator ...Cofactor required for Sp1 transcriptional activation subunit 9 / CRSP complex subunit 9 / Mediator complex subunit 7


Mass: 18836.996 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CZB6
#5: Protein Mediator of RNA polymerase II transcription subunit 8 / Activator-recruited cofactor 32 kDa component / ARC32 / Mediator complex subunit 8


Mass: 20156.684 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9D7W5
#6: Protein Mediator of RNA polymerase II transcription subunit 9 / Mediator complex subunit 9


Mass: 8924.443 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q8VCS6
#7: Protein Mediator of RNA polymerase II transcription subunit 10 / Mediator complex subunit 10


Mass: 15951.878 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CXU0
#8: Protein Mediator of RNA polymerase II transcription subunit 11 / Mediator complex subunit 11


Mass: 12166.723 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9D8C6
#9: Protein Mediator of RNA polymerase II transcription subunit 14 / Cofactor required for Sp1 transcriptional activation subunit 2 / CRSP complex subunit 2 / Mediator ...Cofactor required for Sp1 transcriptional activation subunit 2 / CRSP complex subunit 2 / Mediator complex subunit 14 / Thyroid hormone receptor-associated protein complex 170 kDa component / Trap170


Mass: 168719.531 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: A2ABV5
#10: Protein Mediator of RNA polymerase II transcription subunit 15 / Mediator complex subunit 15 / Positive cofactor 2 glutamine/Q-rich-associated protein / mPcqap


Mass: 18904.717 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q924H2
#11: Protein Mediator of RNA polymerase II transcription subunit 16 / Mediator complex subunit 16 / Thyroid hormone receptor-associated protein 5 / Thyroid hormone ...Mediator complex subunit 16 / Thyroid hormone receptor-associated protein 5 / Thyroid hormone receptor-associated protein complex 95 kDa component / Trap95 / Vitamin D3 receptor-interacting protein complex 92 kDa component / DRIP92


Mass: 91890.484 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q6PGF3
#12: Protein Mediator of RNA polymerase II transcription subunit 17 / Cofactor required for Sp1 transcriptional activation subunit 6 / CRSP complex subunit 6 / Mediator ...Cofactor required for Sp1 transcriptional activation subunit 6 / CRSP complex subunit 6 / Mediator complex subunit 17 / Thyroid hormone receptor-associated protein complex 80 kDa component


Mass: 70669.320 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q8VCD5
#13: Protein Mediator of RNA polymerase II transcription subunit 18 / Mediator complex subunit 18


Mass: 21433.850 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CZ82
#14: Protein Mediator of RNA polymerase II transcription subunit 19 / Mediator complex subunit 19


Mass: 24684.615 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q8C1S0
#15: Protein Mediator of RNA polymerase II transcription subunit 20 / Mediator complex subunit 20 / TRF-proximal protein homolog


Mass: 21660.906 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9R0X0
#16: Protein Mediator of RNA polymerase II transcription subunit 21 / Mediator complex subunit 21 / RNA polymerase II holoenzyme component SRB7 / RNAPII complex component SRB7


Mass: 13619.062 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CQ39
#17: Protein Mediator of RNA polymerase II transcription subunit 22 / Mediator complex subunit 22 / Surfeit locus protein 5 / Surf-5


Mass: 15284.022 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q62276
#18: Protein Mediator of RNA polymerase II transcription subunit 23 / Cofactor required for Sp1 transcriptional activation subunit 3 / CRSP complex subunit 3 / Mediator ...Cofactor required for Sp1 transcriptional activation subunit 3 / CRSP complex subunit 3 / Mediator complex subunit 23 / Protein sur-2 homolog / mSur-2


Mass: 156265.656 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q80YQ2
#19: Protein Mediator of RNA polymerase II transcription subunit 24 / Mediator complex subunit 24 / Thyroid hormone receptor-associated protein 4 / Thyroid hormone ...Mediator complex subunit 24 / Thyroid hormone receptor-associated protein 4 / Thyroid hormone receptor-associated protein complex 100 kDa component / mTRAP100


Mass: 109563.117 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q99K74
#20: Protein Mediator of RNA polymerase II transcription subunit 25 / Mediator complex subunit 25 / mMED25


Mass: 22032.941 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q8VCB2
#21: Protein Mediator of RNA polymerase II transcription subunit 27 / Cofactor required for Sp1 transcriptional activation subunit 8 / CRSP complex subunit 8 / Mediator ...Cofactor required for Sp1 transcriptional activation subunit 8 / CRSP complex subunit 8 / Mediator complex subunit 27


Mass: 33737.773 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9DB40
#22: Protein Mediator of RNA polymerase II transcription subunit 28 / Mediator complex subunit 28


Mass: 13531.168 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q920D3
#23: Protein Mediator of RNA polymerase II transcription subunit 29 / Intersex-like protein / Mediator complex subunit 29


Mass: 14965.154 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9DB91
#24: Protein Mediator of RNA polymerase II transcription subunit 30 / Mediator complex subunit 30 / Thyroid hormone receptor-associated protein 6 / Thyroid hormone ...Mediator complex subunit 30 / Thyroid hormone receptor-associated protein 6 / Thyroid hormone receptor-associated protein complex 25 kDa component / Trap25


Mass: 17873.512 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CQI9
#25: Protein Mediator of RNA polymerase II transcription subunit 31 / Mediator complex subunit 31 / Mediator complex subunit SOH1


Mass: 11545.370 Da / Num. of mol.: 1 / Source method: isolated from a natural source / Source: (natural) Mus musculus (house mouse) / References: UniProt: Q9CXU1

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Experimental details

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Experiment

ExperimentMethod: ELECTRON MICROSCOPY
EM experimentAggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction

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Sample preparation

ComponentName: Mouse mediator complexMediator (coactivator) / Type: COMPLEX / Entity ID: all / Source: NATURAL
Molecular weightValue: 1.0 MDa / Experimental value: NO
Source (natural)Organism: Mus musculus (house mouse)
Buffer solutionpH: 7.9
SpecimenConc.: 0.1 mg/ml / Embedding applied: YES / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
Specimen supportGrid material: COPPER / Grid mesh size: 400 divisions/in. / Grid type: PELCO Ultrathin Carbon with Lacey Carbon
EM embeddingMaterial: ice
VitrificationInstrument: HOMEMADE PLUNGER / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K

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Electron microscopy imaging

Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company
MicroscopyModel: FEI TALOS ARCTICA
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / C2 aperture diameter: 50 µm
Image recordingElectron dose: 22.5 e/Å2 / Film or detector model: GATAN K3 (6k x 4k)

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Processing

CTF correctionType: NONE
3D reconstructionResolution: 4.02 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 217557 / Symmetry type: POINT

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