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- EMDB-8479: Cryo-EM map of the transcriptional Mediator -

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Basic information

Entry
Database: EMDB / ID: EMD-8479
TitleCryo-EM map of the transcriptional Mediator
Map dataSp Mediator
Sample
  • Complex: Mediator complexMediator (coactivator)
    • Protein or peptide: x 16 types
Keywordstranscription / transcriptional / Mediator / Head / Middle / Tail / Srb / RNA polymerase II / Med
Function / homology
Function and homology information


core mediator complex / mediator complex / termination of RNA polymerase II transcription / positive regulation of transcription initiation by RNA polymerase II / transcription initiation at RNA polymerase II promoter / transcription coregulator activity / euchromatin / transcription coactivator activity / RNA polymerase II cis-regulatory region sequence-specific DNA binding / regulation of transcription by RNA polymerase II ...core mediator complex / mediator complex / termination of RNA polymerase II transcription / positive regulation of transcription initiation by RNA polymerase II / transcription initiation at RNA polymerase II promoter / transcription coregulator activity / euchromatin / transcription coactivator activity / RNA polymerase II cis-regulatory region sequence-specific DNA binding / regulation of transcription by RNA polymerase II / nucleus / cytoplasm
Similarity search - Function
Mediator complex, subunit Med27 / Mediator complex subunit 27 / Mediator complex, subunit Med6, fungi / Mediator complex, subunit Med7 superfmaily / 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 ...Mediator complex, subunit Med27 / Mediator complex subunit 27 / Mediator complex, subunit Med6, fungi / Mediator complex, subunit Med7 superfmaily / 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 / 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
Similarity search - Domain/homology
Uncharacterized protein C24C9.04 / Mediator of RNA polymerase II transcription subunit 22 / Mediator of RNA polymerase II transcription subunit 18 / Mediator of RNA polymerase II transcription subunit 7 / Mediator of RNA polymerase II transcription subunit 21 / Mediator of RNA polymerase II transcription subunit 8 / Mediator of RNA polymerase II transcription subunit 17 / Mediator of RNA polymerase II transcription subunit 20 / Mediator of RNA polymerase II transcription subunit 27 / Mediator of RNA polymerase II transcription subunit 11 ...Uncharacterized protein C24C9.04 / Mediator of RNA polymerase II transcription subunit 22 / Mediator of RNA polymerase II transcription subunit 18 / Mediator of RNA polymerase II transcription subunit 7 / Mediator of RNA polymerase II transcription subunit 21 / Mediator of RNA polymerase II transcription subunit 8 / Mediator of RNA polymerase II transcription subunit 17 / Mediator of RNA polymerase II transcription subunit 20 / Mediator of RNA polymerase II transcription subunit 27 / Mediator of RNA polymerase II transcription subunit 11 / Mediator of RNA polymerase II transcription subunit 14 / Mediator of RNA polymerase II transcription subunit 6 / Mediator of RNA polymerase II transcription subunit 31
Similarity search - Component
Biological speciesSchizosaccharomyces pombe (fission yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.4 Å
AuthorsTsai K-L / Yu X
CitationJournal: Nature / Year: 2017
Title: Mediator structure and rearrangements required for holoenzyme formation.
Authors: Kuang-Lei Tsai / Xiaodi Yu / Sneha Gopalan / Ti-Chun Chao / Ying Zhang / Laurence Florens / Michael P Washburn / Kenji Murakami / Ronald C Conaway / Joan W Conaway / Francisco J Asturias /
Abstract: The conserved Mediator co-activator complex has an essential role in the regulation of RNA polymerase II transcription in all eukaryotes. Understanding the structure and interactions of Mediator is ...The conserved Mediator co-activator complex has an essential role in the regulation of RNA polymerase II transcription in all eukaryotes. Understanding the structure and interactions of Mediator is crucial for determining how the complex influences transcription initiation and conveys regulatory information to the basal transcription machinery. Here we present a 4.4 Å resolution cryo-electron microscopy map of Schizosaccharomyces pombe Mediator in which conserved Mediator subunits are individually resolved. The essential Med14 subunit works as a central backbone that connects the Mediator head, middle and tail modules. Comparison with a 7.8 Å resolution cryo-electron microscopy map of a Mediator-RNA polymerase II holoenzyme reveals that changes in the structure of Med14 facilitate a large-scale Mediator rearrangement that is essential for holoenzyme formation. Our study suggests that access to different conformations and crosstalk between structural elements are essential for the Mediator regulation mechanism, and could explain the capacity of the complex to integrate multiple regulatory signals.
History
DepositionNov 26, 2016-
Header (metadata) releaseMar 8, 2017-
Map releaseMar 8, 2017-
UpdateMar 13, 2024-
Current statusMar 13, 2024Processing site: RCSB / Status: Released

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Structure visualization

Movie
  • Surface view with section colored by density value
  • Surface level: 0.016
  • Imaged by UCSF Chimera
  • Download
  • Surface view colored by radius
  • Surface level: 0.016
  • Imaged by UCSF Chimera
  • Download
  • Surface view with fitted model
  • Atomic models: PDB-5u0p
  • Surface level: 0.016
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerEM map:
SurfViewMolmilJmol/JSmol
Supplemental images

emd_8479.png

Downloads & links

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Map

FileDownload / File: emd_8479.map.gz / Format: CCP4 / Size: 166.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationSp Mediator
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.31 Å/pix.
x 352 pix.
= 461.12 Å		1.31 Å/pix.
x 352 pix.
= 461.12 Å		1.31 Å/pix.
x 352 pix.
= 461.12 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.31 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.016 / Movie #1: 0.016
Minimum - Maximum-0.017003836 - 0.07525921
Average (Standard dev.)-0.000033825316 (±0.0022060836)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions352352352
Spacing352352352
CellA=B=C: 461.12 Å
α=β=γ: 90.0 °

CCP4 map header:

modeImage stored as Reals
Å/pix. X/Y/Z1.311.311.31
M x/y/z352352352
origin x/y/z0.0000.0000.000
length x/y/z461.120461.120461.120
α/β/γ90.00090.00090.000
MAP C/R/S123
start NC/NR/NS000
NC/NR/NS352352352
D min/max/mean-0.0170.075-0.000

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Supplemental data

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Mask #1

Fileemd_8479_msk_1.map
Projections & Slices
AxesZYX

Projections

Slices (1/2)
Density Histograms

Histogram

Histogram (log scale)

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Sample components

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Entire : Mediator complex

EntireName: Mediator complexMediator (coactivator)
Components
  • Complex: Mediator complexMediator (coactivator)
    • Protein or peptide: Mediator complex subunit 14Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 6Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 8Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 17Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 18Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 20Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 11Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 22Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 4Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 7Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 21Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 31Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 9Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 27Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 19Mediator (coactivator)
    • Protein or peptide: Mediator complex subunit 10Mediator (coactivator)

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Supramolecule #1: Mediator complex

SupramoleculeName: Mediator complex / type: complex / ID: 1 / Parent: 0 / Macromolecule list: all
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)

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Macromolecule #1: Mediator complex subunit 14

MacromoleculeName: Mediator complex subunit 14 / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 105.382461 KDa
SequenceString: MEPPAIPHIT EGFYPLPEIV ETFSHHVLQE LVSLAEVLPS MSNVEKKKKI LDWLLRSRAF TMRLLVLARW VHLSPSVHRC IDVVAFLQG QKFCFQNLVH VLQDIRYQLS FARLRNSDLV TALDILSTGT SLRLANAPTS KLYMLSESPL STKQILQTLH A LNMLIRIR ...String:
MEPPAIPHIT EGFYPLPEIV ETFSHHVLQE LVSLAEVLPS MSNVEKKKKI LDWLLRSRAF TMRLLVLARW VHLSPSVHRC IDVVAFLQG QKFCFQNLVH VLQDIRYQLS FARLRNSDLV TALDILSTGT SLRLANAPTS KLYMLSESPL STKQILQTLH A LNMLIRIR LSLYEIIPTP FQHFTIANGR CTFTVPNEFS VSLTTNSQDP KSTGISFQWI VVDFQFHLPD FSSTPAKYRV FI ELHLNEE IAAAFVLQKP ILPLIYNILH KFCLYQRLNL LSQQTFQLSR ESWLGHLRGV YDEKPPRLRL YYWPQLNVKK EGK PGKIGH YIHIFVNTQP ISAFERTLSS KRSSCEYDHF LLLVEWHHDG IVEHVPLDDH MDAQHLLLLI TQKHAQLILE QIRK ELHPN IFSEHVGGGL KIHVFDNEII VKVNSVTGRL VLSSSASPLS PPRHLRAAEK NIALNTQPPA QILNRLYFFC IQTQL LEVA QCAELHAVQG YYSFPYLTFS KGKWRKDGDS LWVLAYNVES NSWSVRLLNA AGQTLYTQDV HTTKGTLSIE SFSRLS YLL EVQILLFNVQ TACQARGMPF EYLPIPPKAL IEDDFTTYVQ TGCLCIMMPS SNEDMLPVVF VRAHDGQLIF DSRIKGK LP YQSETETEKN CYIDWRTGRI TIRVQNFSSF EKTWIGLLKL VALSKTSAFN VDCITLKHVD FTYLDDEKFR ATIHDDNT F TLHFFNRHSP FHLISQFLQD TFSDGPSAIQ PLRVIMDRTR GVLVAQELGY VVLARSLRQY RIILSKNHGI QVLLNRHGC ILQDLSYLSA DSRYLEGTQT LTSQWEPCSW LNTVWEGDLG DDELNGQIEA APEMHLIKMN KTADLTAILK RILAISRKK (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)

UniProtKB: Mediator of RNA polymerase II transcription subunit 14

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Macromolecule #2: Mediator complex subunit 6

MacromoleculeName: Mediator complex subunit 6 / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 24.928947 KDa
SequenceString: MASGAPPSVD LTSIQWRMPE WVQSMGGLRT ENVLEYFSQS PFYSHKSNNE MLKMQSQFNA LDLGDLNSQL KRLTGIQFVI IHERPPFLW VIQKQNRLNE NEVKPLTVYF VCNENIYMAP NAYTLLATRM LNATYCFQKA LTKIEKFPQY NPQEGYTYPK L SNDNLEVD ...String:
MASGAPPSVD LTSIQWRMPE WVQSMGGLRT ENVLEYFSQS PFYSHKSNNE MLKMQSQFNA LDLGDLNSQL KRLTGIQFVI IHERPPFLW VIQKQNRLNE NEVKPLTVYF VCNENIYMAP NAYTLLATRM LNATYCFQKA LTKIEKFPQY NPQEGYTYPK L SNDNLEVD HSNTNEPADE NKNQSIENAD YSFSPEDFSV VRAFMQSLHS SKEAPDVK

UniProtKB: Mediator of RNA polymerase II transcription subunit 6

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Macromolecule #3: Mediator complex subunit 8

MacromoleculeName: Mediator complex subunit 8 / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 23.360068 KDa
SequenceString: MEDISTEKTV ESLEAIRHRI AQIVQSLTHF LAILHQSESL SPWPTIHKNF NILLSQIHSL SNNLAAHSHT LQTTSIYPSL EFPVKEQEP LLTTLLRTKA LPEVEEWEAN TLQEYEASIS SQPKKKEAND AYQKDQLWDQ ARIIFMEERE NYSWFDFVTR R QESEGEFV ...String:
MEDISTEKTV ESLEAIRHRI AQIVQSLTHF LAILHQSESL SPWPTIHKNF NILLSQIHSL SNNLAAHSHT LQTTSIYPSL EFPVKEQEP LLTTLLRTKA LPEVEEWEAN TLQEYEASIS SQPKKKEAND AYQKDQLWDQ ARIIFMEERE NYSWFDFVTR R QESEGEFV SQRQLEIDRA TEEQNANQML TDILSFMKSG KR

UniProtKB: Mediator of RNA polymerase II transcription subunit 8

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Macromolecule #4: Mediator complex subunit 17

MacromoleculeName: Mediator complex subunit 17 / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 62.55141 KDa
SequenceString: MAEEANKDAD ISSLSLSLDP EIIGGQNNFL ENNLQQIFQK IIQERGPFRD LKEEDLQKEL QKESIKDESS AKSSETENVL EFATLDSKR NVNDTEVESM DSQAYKKELI EQIMIAQTEC SLALDMTSLL LSKFKENSIE TISPFLKSTV PPSSLQFSRS Q PPESKESD ...String:
MAEEANKDAD ISSLSLSLDP EIIGGQNNFL ENNLQQIFQK IIQERGPFRD LKEEDLQKEL QKESIKDESS AKSSETENVL EFATLDSKR NVNDTEVESM DSQAYKKELI EQIMIAQTEC SLALDMTSLL LSKFKENSIE TISPFLKSTV PPSSLQFSRS Q PPESKESD ATLAKCWKEK SLTSSCKFLF EAKERLTSVV ETEHEYYTEL VKVKEASWPL FNSQGSNHLS VQYSCLGGIS LG LGLIRMK PESKSFEVQS SLLYSQAALK ISILNKDRDE IGSSTWSWPS QNCNSVLLKD IYKLQEILFE MDIWNSLLQE AQS CGNQGV NFTGDEILVP ISDDHVVRIT LETSSKNTES GFTEDKKSNE DTSTNFVTIK QEKELLKCLC DTLNAIAHIL FLKH CRKSD RRSQQPELYM AIDANAPLIL RPLIFYYNLN QESLEFQRWL KQRDISFKFM PNYPWEKAKD FLELENSLSI NRLSI SWRI MVSNFEPAIF IQHTPTLHGT DKSVWRCKDQ YSSNQFSSLK NVCQYIEHHI NSLSRRSKKT E

UniProtKB: Mediator of RNA polymerase II transcription subunit 17

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Macromolecule #5: Mediator complex subunit 18

MacromoleculeName: Mediator complex subunit 18 / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 24.056377 KDa
SequenceString: MQELYLLGVV PSRRFEAVVN SLSKTLDGPK TILEFWVVYR PKDVPPNLPR QPDSWLRLCS NIESHDETDT EWSKNTQWSM YLEGNSEPK REDKCGIRPV NRAKLTNGSV TEFVEKMGYE FSHEYIIQGL EYFFFDTTVR IYQTLIPSQQ RSIKPPFHPM N EEQPWILH ...String:
MQELYLLGVV PSRRFEAVVN SLSKTLDGPK TILEFWVVYR PKDVPPNLPR QPDSWLRLCS NIESHDETDT EWSKNTQWSM YLEGNSEPK REDKCGIRPV NRAKLTNGSV TEFVEKMGYE FSHEYIIQGL EYFFFDTTVR IYQTLIPSQQ RSIKPPFHPM N EEQPWILH VYTHVADASN QVAMAKAEAN LTKVKTLLSA FCDLKNVRL

UniProtKB: Mediator of RNA polymerase II transcription subunit 18

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Macromolecule #6: Mediator complex subunit 20

MacromoleculeName: Mediator complex subunit 20 / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 22.374775 KDa
SequenceString: MPVHGVIYYS SPSMATFLSP AQDNLVRTYF AQHLKKWVVQ YKLYRNAVTP KTLEFLKQNI NPSMLACVDE ATMIDAEPEL EDIIVRTKL WNFRQSFTIE GSIYEVGSFK VAIANVLQKS VWKGILFHVT YDGTESVDLA RPIIQEFFLK CFLQNNKSVT P VYESFFNQ ...String:
MPVHGVIYYS SPSMATFLSP AQDNLVRTYF AQHLKKWVVQ YKLYRNAVTP KTLEFLKQNI NPSMLACVDE ATMIDAEPEL EDIIVRTKL WNFRQSFTIE GSIYEVGSFK VAIANVLQKS VWKGILFHVT YDGTESVDLA RPIIQEFFLK CFLQNNKSVT P VYESFFNQ PRHSLDSKLL LQLFKQRIDT VSQRT

UniProtKB: Mediator of RNA polymerase II transcription subunit 20

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Macromolecule #7: Mediator complex subunit 11

MacromoleculeName: Mediator complex subunit 11 / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 13.1188 KDa
SequenceString:
MTNSDDDLFS EKSTSSDTQQ VQNILELEAK IPDILSSAGK CIEAIQLNNS LEDFRKYSKE FLETVEFIST GLRRQALELE KAEVPVVSL QPKKRYASTP LSNLIFDQSS KLIPKYC

UniProtKB: Mediator of RNA polymerase II transcription subunit 11

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Macromolecule #8: Mediator complex subunit 22

MacromoleculeName: Mediator complex subunit 22 / type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 15.396165 KDa
SequenceString:
MSSDSFQRQL VQRTNTLNSS IDNATLTILS RFQDILDIAI NEGKDKYTVA PEVYQIECHT VSMVRAVEQL LDVSRQIKSY WLTNSLSTS FPTVDYSEPD LEKVKRTLTK LQNHLLEVSL IEPEASETTE APTVSDT

UniProtKB: Mediator of RNA polymerase II transcription subunit 22

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Macromolecule #9: Mediator complex subunit 4

MacromoleculeName: Mediator complex subunit 4 / type: protein_or_peptide / ID: 9 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 20.35801 KDa
SequenceString: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) ...String:
(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)

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Macromolecule #10: Mediator complex subunit 7

MacromoleculeName: Mediator complex subunit 7 / type: protein_or_peptide / ID: 10 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 43.573094 KDa
SequenceString: MEPNEGVQLF SAFPPPPPYY KLFTRENIEK VISNMEKEAK HEDDANTLQP KTEEEIESLA KLFKKPSCLT SGTYQMFGDT WRLDEAIPS LKEFGIPELY KDIKDGEDEI EVVEYDPKSN AIVGTSFTRV HDYDKNSPIE NEKILEDDQH TAMKEKDNES D KTMKDVEE ...String:
MEPNEGVQLF SAFPPPPPYY KLFTRENIEK VISNMEKEAK HEDDANTLQP KTEEEIESLA KLFKKPSCLT SGTYQMFGDT WRLDEAIPS LKEFGIPELY KDIKDGEDEI EVVEYDPKSN AIVGTSFTRV HDYDKNSPIE NEKILEDDQH TAMKEKDNES D KTMKDVEE KTEPSLKKEE EDIQMKEPLD SQDTGAVSAS SVNEGFRADQ KSKDGETSDL IKIPRRAYEL RFLSRSLMLN FL ELLGIMA KAPEQFPSKV ENIRVLLLNL HHLINDYRPH QSRESLIMLL EKQLKHEESQ VELLRTHNRQ MTETLEKYKS LDF NMEKEG DVIQQLKSSI KKPLSGAEDE QKSRSMFSKN DEKLKKSLEL MEDVIKRDLS

UniProtKB: Mediator of RNA polymerase II transcription subunit 7

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Macromolecule #11: Mediator complex subunit 21

MacromoleculeName: Mediator complex subunit 21 / type: protein_or_peptide / ID: 11 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 15.821031 KDa
SequenceString:
MACRCTQLQD TIDEVATQFY SSIHYLSSHH DFVPLPGQEK VSDSKVNPIS AEELQFAQRD LAKDLVTKFM QIDTLINQLP GISTAPKHQ LEKIKKLQNS IEEKQLERKS LESENEDLKL QLAKRIETFG RLSCVLFQP

UniProtKB: Mediator of RNA polymerase II transcription subunit 21

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Macromolecule #12: Mediator complex subunit 31

MacromoleculeName: Mediator complex subunit 31 / type: protein_or_peptide / ID: 12 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 16.913975 KDa
SequenceString:
METKWLLSKV PDDKSRFEIE LEFVQMLSNP WYLNFLAQHK YFEDEAFLQY LEYMEYWREP EYVKFIIYPT CLHMLTLLKN PQFRNDISR ADLSKQVNDE IYYEWLGKGL QQYGSADDAT LSQPQQEEDE KKVDVKKENE

UniProtKB: Mediator of RNA polymerase II transcription subunit 31

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Macromolecule #13: Mediator complex subunit 9

MacromoleculeName: Mediator complex subunit 9 / type: protein_or_peptide / ID: 13 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 13.810964 KDa
SequenceString:
MDPAHHIKDT SAELLSQQDF SILQSRLLEF LASQEARETV TASKELTLLR QGIRQLKEKV SKMEPEEMTV KEKKSIIEIL KARIALKKA FLKMALSESN DTVVKKEDVR ESPVDTFMEN AT

UniProtKB: Uncharacterized protein C24C9.04

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Macromolecule #14: Mediator complex subunit 27

MacromoleculeName: Mediator complex subunit 27 / type: protein_or_peptide / ID: 14 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 31.132693 KDa
SequenceString: MSLEEQRTRD ELKHKLLDLN QLHEQLAELR TICPSLLKLL HPETGTSRKF EKSAQEAIEK VNSFYTHLKS SQNVFDYAEK SLQADSSNL LPTYLYNSED LSNDTENNET KSINGKSALD LKEPHHSELH DNDNFQNSDI NIESFKGDIE ASGSILTTHE N KSFTLKLA ...String:
MSLEEQRTRD ELKHKLLDLN QLHEQLAELR TICPSLLKLL HPETGTSRKF EKSAQEAIEK VNSFYTHLKS SQNVFDYAEK SLQADSSNL LPTYLYNSED LSNDTENNET KSINGKSALD LKEPHHSELH DNDNFQNSDI NIESFKGDIE ASGSILTTHE N KSFTLKLA NELEFIFFHD TRGKFSVYCS SSKDDAITFS INRNNNFLGN LWSLMPKILD YQHLYSKPCD FCKSLISPVY LE LPSVRRN ANSTVKPTSK DILALHAECV PAQSDL

UniProtKB: Mediator of RNA polymerase II transcription subunit 27

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Macromolecule #15: Mediator complex subunit 19

MacromoleculeName: Mediator complex subunit 19 / type: protein_or_peptide / ID: 15 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 11.847596 KDa
SequenceString: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) ...String:
(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)

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Macromolecule #16: Mediator complex subunit 10

MacromoleculeName: Mediator complex subunit 10 / type: protein_or_peptide / ID: 16 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Schizosaccharomyces pombe (fission yeast)
Molecular weightTheoretical: 11.251861 KDa
SequenceString: (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) ...String:
(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK) (UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)(UNK) (UNK)(UNK)(UNK)(UNK)

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Experimental details

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Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

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Sample preparation

Concentration1 mg/mL
BufferpH: 7.4
Details: 20 mM HEPES, pH 7.4, 200 mM potassium acetate, 5 mM b-Mercaptoethanol, 0.01% NP-40
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsC2 aperture diameter: 100.0 µm / Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Cs: 2.7 mm / Nominal defocus max: 4.0 µm / Nominal defocus min: 1.0 µm / Nominal magnification: 22500
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 11181 / Average exposure time: 7.0 sec. / Average electron dose: 9.6 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Startup modelType of model: NONE
Initial angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 1.4)
Final angle assignmentType: ANGULAR RECONSTITUTION / Software - Name: RELION (ver. 1.4)
Final reconstructionApplied symmetry - Point group: C1 (asymmetric) / Resolution.type: BY AUTHOR / Resolution: 4.4 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 1.4) / Number images used: 42484
FSC plot (resolution estimation)

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