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- EMDB-29071: TFIIIA-TFIIIC-Brf1-TBP complex bound to 5S rRNA gene -

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Basic information

Entry
Database: EMDB / ID: EMD-29071
TitleTFIIIA-TFIIIC-Brf1-TBP complex bound to 5S rRNA gene
Map dataMain map postprocessed in Relion
Sample
  • Complex: TFIIIA-TFIIIC-Brf1-TBP complex bound to 5SrRNA gene
    • Protein or peptide: x 8 types
    • DNA: x 2 types
  • Ligand: x 1 types
Keywordstranscription factor / TRANSCRIPTION / TRANSCRIPTION-DNA complex
Function / homology
Function and homology information


RNA polymerase II sequence-specific DNA-binding transcription factor recruiting activity / 5S class rRNA transcription by RNA polymerase III / transcription factor TFIIIC complex / RNA polymerase III core binding / RNA polymerase III type 1 promoter sequence-specific DNA binding / RNA polymerase III type 2 promoter sequence-specific DNA binding / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / DNA-templated transcription open complex formation / transcription factor TFIIIB complex ...RNA polymerase II sequence-specific DNA-binding transcription factor recruiting activity / 5S class rRNA transcription by RNA polymerase III / transcription factor TFIIIC complex / RNA polymerase III core binding / RNA polymerase III type 1 promoter sequence-specific DNA binding / RNA polymerase III type 2 promoter sequence-specific DNA binding / TFIIA-class transcription factor complex binding / RNA polymerase III transcription regulatory region sequence-specific DNA binding / DNA-templated transcription open complex formation / transcription factor TFIIIB complex / RNA polymerase III preinitiation complex assembly / TFIIIC-class transcription factor complex binding / RNA polymerase III type 3 promoter sequence-specific DNA binding / RNA polymerase I general transcription initiation factor binding / RNA polymerase III general transcription initiation factor activity / regulation of transcription by RNA polymerase III / transcription factor TFIIA complex / RNA polymerase I preinitiation complex assembly / transcription preinitiation complex / DNA binding, bending / RNA Polymerase III Transcription Initiation From Type 2 Promoter / RNA polymerase II transcribes snRNA genes / RNA Polymerase II Promoter Escape / RNA Polymerase II Transcription Pre-Initiation And Promoter Opening / RNA Polymerase II Transcription Initiation / RNA Polymerase II Transcription Initiation And Promoter Clearance / RNA Polymerase II Pre-transcription Events / RNA Polymerase I Promoter Escape / transcription initiation at RNA polymerase III promoter / nucleolar large rRNA transcription by RNA polymerase I / phosphatase activity / Estrogen-dependent gene expression / transcription by RNA polymerase III / transcription factor TFIID complex / RNA polymerase II general transcription initiation factor activity / RNA polymerase II core promoter sequence-specific DNA binding / RNA polymerase II preinitiation complex assembly / protein localization to chromatin / TBP-class protein binding / DNA-templated transcription initiation / disordered domain specific binding / DNA-binding transcription factor binding / RNA polymerase II-specific DNA-binding transcription factor binding / transcription regulator complex / DNA-binding transcription factor activity, RNA polymerase II-specific / RNA polymerase II cis-regulatory region sequence-specific DNA binding / negative regulation of DNA-templated transcription / chromatin binding / regulation of DNA-templated transcription / positive regulation of transcription by RNA polymerase II / protein-containing complex / mitochondrion / DNA binding / RNA binding / nucleoplasm / nucleus / metal ion binding / cytoplasm
Similarity search - Function
Transcription factor tau subunit sfc3/Tfc3, C-terminal / : / Family of unknown function (DUF6581) / Transcription factor tau 138 kDa subunit, extended winged helix / Transcription factor TFIIIC subunit Tfc7/tau55 / Transcription factor IIIC subunit 5, HTH domain / Transcription factor TFIIIC, triple barrel domain / Transcription factor Tfc4/TFIIIC-102/Sfc4 / Transcription factor IIIC subunit Tfc1/Sfc1 / Transcription factor IIIC subunit Tfc1/Sfc1, triple barrel domain ...Transcription factor tau subunit sfc3/Tfc3, C-terminal / : / Family of unknown function (DUF6581) / Transcription factor tau 138 kDa subunit, extended winged helix / Transcription factor TFIIIC subunit Tfc7/tau55 / Transcription factor IIIC subunit 5, HTH domain / Transcription factor TFIIIC, triple barrel domain / Transcription factor Tfc4/TFIIIC-102/Sfc4 / Transcription factor IIIC subunit Tfc1/Sfc1 / Transcription factor IIIC subunit Tfc1/Sfc1, triple barrel domain / TFIIIC, subcomplex tauA subunit Sfc1, triple barrel domain superfamily / RNA polymerase III transcription factor (TF)IIIC subunit HTH domain / TFIIIC subunit triple barrel domain / Tau95 Triple barrel domain / Transcription factor IIIC, putative zinc-finger / Putative zinc-finger of transcription factor IIIC complex / B-block binding subunit of TFIIIC / Tfc3, extended winged-helix domain / Transcription facto Tfc3-like / B-block binding subunit of TFIIIC / Brf1, TBP-binding domain / Brf1-like TBP-binding domain / C2H2-type zinc finger / Phosphoglycerate mutase family / Histidine phosphatase superfamily, clade-1 / Histidine phosphatase superfamily (branch 1) / Transcription factor TFIIB, cyclin-like domain / Transcription factor TFIIB, conserved site / Transcription factor TFIIB repeat / Transcription factor TFIIB repeat signature. / Transcription factor TFIIB / Zinc finger TFIIB-type profile. / Zinc finger, TFIIB-type / TFIIB zinc-binding / TATA-box binding protein, eukaryotic / TATA-box binding protein / TATA-box binding protein, conserved site / Transcription factor TFIID (or TATA-binding protein, TBP) / Transcription factor TFIID repeat signature. / Histidine phosphatase superfamily / Tetratricopeptide repeat / Tetratricopeptide repeat / TBP domain superfamily / Zinc finger, C2H2 type / Cyclin-like / domain present in cyclins, TFIIB and Retinoblastoma / Cyclin-like superfamily / zinc finger / Zinc finger C2H2 type domain profile. / TPR repeat region circular profile. / TPR repeat profile. / Zinc finger C2H2 superfamily / Zinc finger C2H2 type domain signature. / Tetratricopeptide repeats / Tetratricopeptide repeat / Zinc finger C2H2-type / Tetratricopeptide-like helical domain superfamily / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
Similarity search - Domain/homology
TATA-box-binding protein / Transcription factor IIIB 70 kDa subunit / Transcription factor tau 95 kDa subunit / Transcription factor tau 131 kDa subunit / Transcription factor tau 138 kDa subunit / Transcription factor IIIA / Transcription factor tau 91 kDa subunit / Transcription factor tau 60 kDa subunit / Transcription factor tau 55 kDa subunit
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast) / Saccharomyces cerevisiae S288C (yeast)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.8 Å
AuthorsTalyzina A / He Y
Funding support United States, 3 items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS) United States
American Cancer Society United States
National Institutes of Health/National Cancer Institute (NIH/NCI) United States
CitationJournal: bioRxiv / Year: 2023
Title: Structural basis of TFIIIC-dependent RNA Polymerase III transcription initiation.
Authors: Anna Talyzina / Yan Han / Chiranjib Banerjee / Susan Fishbain / Alexis Reyes / Reza Vafabakhsh / Yuan He /
Abstract: RNA Polymerase III (Pol III) is responsible for transcribing 5S ribosomal RNA (5S rRNA), tRNAs, and other short non-coding RNAs. Its recruitment to the 5S rRNA promoter requires transcription factors ...RNA Polymerase III (Pol III) is responsible for transcribing 5S ribosomal RNA (5S rRNA), tRNAs, and other short non-coding RNAs. Its recruitment to the 5S rRNA promoter requires transcription factors TFIIIA, TFIIIC, and TFIIIB. Here we use cryo-electron microscopy to visualize the complex of TFIIIA and TFIIIC bound to the promoter. Brf1-TBP binding further stabilizes the DNA, resulting in the full-length 5S rRNA gene wrapping around the complex. Our smFRET study reveals that the DNA undergoes both sharp bending and partial dissociation on a slow timescale, consistent with the model predicted from our cryo-EM results. Our findings provide new insights into the mechanism of how the transcription initiation complex assembles on the 5S rRNA promoter, a crucial step in Pol III transcription regulation.
History
DepositionDec 11, 2022-
Header (metadata) releaseJun 21, 2023-
Map releaseJun 21, 2023-
UpdateAug 16, 2023-
Current statusAug 16, 2023Processing site: RCSB / Status: Released

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Structure visualization

Supplemental images

emd_29071.png

Downloads & links

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Map

FileDownload / File: emd_29071.map.gz / Format: CCP4 / Size: 216 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationMain map postprocessed in Relion
Voxel sizeX=Y=Z: 1.056 Å
Density
Contour LevelBy AUTHOR: 0.016
Minimum - Maximum-0.06010179 - 0.1019568
Average (Standard dev.)0.00010582097 (±0.0025918644)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions384384384
Spacing384384384
CellA=B=C: 405.504 Å
α=β=γ: 90.0 °

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Supplemental data

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Additional map: Main map postprocessed with deepEMhancer

Fileemd_29071_additional_1.map
AnnotationMain map postprocessed with deepEMhancer
Projections & Slices
AxesZYX

Projections

Slices (1/2)
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Histogram

Histogram (log scale)

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Additional map: Focused refinement of Brf1-TBP-tau131-DNA, postprocessed with deepEMhancer...

Fileemd_29071_additional_2.map
AnnotationFocused refinement of Brf1-TBP-tau131-DNA, postprocessed with deepEMhancer
Projections & Slices
AxesZYX

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Slices (1/2)
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Histogram

Histogram (log scale)

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Half map: Half1 of main map

Fileemd_29071_half_map_1.map
AnnotationHalf1 of main map
Projections & Slices
AxesZYX

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Histogram (log scale)

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Half map: Half2 of main map

Fileemd_29071_half_map_2.map
AnnotationHalf2 of main map
Projections & Slices
AxesZYX

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Sample components

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Entire : TFIIIA-TFIIIC-Brf1-TBP complex bound to 5SrRNA gene

EntireName: TFIIIA-TFIIIC-Brf1-TBP complex bound to 5SrRNA gene
Components
  • Complex: TFIIIA-TFIIIC-Brf1-TBP complex bound to 5SrRNA gene
    • Protein or peptide: Transcription factor IIIA
    • Protein or peptide: Transcription factor tau 138 kDa subunit
    • Protein or peptide: Transcription factor tau 131 kDa subunit
    • Protein or peptide: Transcription factor tau 95 kDa subunit
    • Protein or peptide: Transcription factor tau 91 kDa subunit
    • Protein or peptide: Transcription factor tau 60 kDa subunit
    • Protein or peptide: Transcription factor tau 55 kDa subunit
    • Protein or peptide: Transcription factor IIIB 70 kDa subunit,TATA-box-binding protein
    • DNA: DNA (151-MER)
    • DNA: DNA (151-MER)
  • Ligand: ZINC ION

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Supramolecule #1: TFIIIA-TFIIIC-Brf1-TBP complex bound to 5SrRNA gene

SupramoleculeName: TFIIIA-TFIIIC-Brf1-TBP complex bound to 5SrRNA gene / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#10
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 810 kDa/nm

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Macromolecule #1: Transcription factor IIIA

MacromoleculeName: Transcription factor IIIA / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 52.522516 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MGGEVLNNEG MPLAELKQET IPISRSESSE SLNSLTSTRS SSSNRPKTYF CDYDGCDKAF TRPSILTEHQ LSVHQGLRAF QCDKCAKSF VKKSHLERHL YTHSDTKPFQ CSYCGKGVTT RQQLKRHEVT HTKSFICPEE GCNLRFYKHP QLRAHILSVH L HKLTCPHC ...String:
MGGEVLNNEG MPLAELKQET IPISRSESSE SLNSLTSTRS SSSNRPKTYF CDYDGCDKAF TRPSILTEHQ LSVHQGLRAF QCDKCAKSF VKKSHLERHL YTHSDTKPFQ CSYCGKGVTT RQQLKRHEVT HTKSFICPEE GCNLRFYKHP QLRAHILSVH L HKLTCPHC NKSFQRPYRL RNHISKHHDP EVENPYQCTF AGCCKEFRIW SQLQSHIKND HPKLKCPICS KPCVGENGLQ MH MIIHDDS LVTKNWKCHI CPDMSFSRKH DLLTHYGSIH TEEDIPLELK YKISDIQQLV QDHGVQLGNS KHSNEQDEEK ISN RLRKRR KLTENNNVEF LQNEVDLEKR LESGENGLNL LLNTVGRKYR CFYNNCSRTF KTKEKYEKHI DKHKVHELKL KILQ EKEEN KTLVDQNHKE PFIIQKETQS AGDKADPNSS SVDKLAAALE HHHHHH

UniProtKB: Transcription factor IIIA

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Macromolecule #2: Transcription factor tau 138 kDa subunit

MacromoleculeName: Transcription factor tau 138 kDa subunit / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 132.313062 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MVLTIYPDEL VQIVSDKIAS NKGKITLNQL WDISGKYFDL SDKKVKQFVL SCVILKKDIE VYCDGAITTK NVTDIIGDAN HSYSVGITE DSLWTLLTGY TKKESTIGNS AFELLLEVAK SGEKGINTMD LAQVTGQDPR SVTGRIKKIN HLLTSSQLIY K GHVVKQLK ...String:
MVLTIYPDEL VQIVSDKIAS NKGKITLNQL WDISGKYFDL SDKKVKQFVL SCVILKKDIE VYCDGAITTK NVTDIIGDAN HSYSVGITE DSLWTLLTGY TKKESTIGNS AFELLLEVAK SGEKGINTMD LAQVTGQDPR SVTGRIKKIN HLLTSSQLIY K GHVVKQLK LKKFSHDGVD SNPYINIRDH LATIVEVVKR SKNGIRQIID LKRELKFDKE KRLSKAFIAA IAWLDEKEYL KK VLVVSPK NPAIKIRCVK YVKDIPDSKG SPSFEYDSNS ADEDSVSDSK AAFEDEDLVE GLDNFNATDL LQNQGLVMEE KED AVKNEV LLNRFYPLQN QTYDIADKSG LKGISTMDVV NRITGKEFQR AFTKSSEYYL ESVDKQKENT GGYRLFRIYD FEGK KKFFR LFTAQNFQKL TNAEDEISVP KGFDELGKSR TDLKTLNEDN FVALNNTVRF TTDSDGQDIF FWHGELKIPP NSKKT PNKN KRKRQVKNST NASVAGNISN PKRIKLEQHV STAQEPKSAE DSPSSNGGTV VKGKVVNFGG FSARSLRSLQ RQRAIL KVM NTIGGVAYLR EQFYESVSKY MGSTTTLDKK TVRGDVDLMV ESEKLGARTE PVSGRKIIFL PTVGEDAIQR YILKEKD SK KATFTDVIHD TEIYFFDQTE KNRFHRGKKS VERIRKFQNR QKNAKIKASD DAISKKSTSV NVSDGKIKRR DKKVSAGR T TVVVENTKED KTVYHAGTKD GVQALIRAVV VTKSIKNEIM WDKITKLFPN NSLDNLKKKW TARRVRMGHS GWRAYVDKW KKMLVLAIKS EKISLRDVEE LDLIKLLDIW TSFDEKEIKR PLFLYKNYEE NRKKFTLVRD DTLTHSGNDL AMSSMIQREI SSLKKTYTR KISASTKDLS KSQSDDYIRT VIRSILIESP STTRNEIEAL KNVGNESIDN VIMDMAKEKQ IYLHGSKLEC T DTLPDILE NRGNYKDFGV AFQYRCKVNE LLEAGNAIVI NQEPSDISSW VLIDLISGEL LNMDVIPMVR NVRPLTYTSR RF EIRTLTP PLIIYANSQT KLNTARKSAV KVPLGKPFSR LWVNGSGSIR PNIWKQVVTM VVNEIIFHPG ITLSRLQSRC REV LSLHEI SEICKWLLER QVLITTDFDG YWVNHNWYSI YEST

UniProtKB: Transcription factor tau 138 kDa subunit

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Macromolecule #3: Transcription factor tau 131 kDa subunit

MacromoleculeName: Transcription factor tau 131 kDa subunit / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 120.388609 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MAAGKLKKEQ QNQSAERESA DTGKVNDEDE EHLYGNIDDY KHLIQDEEYD DEDVPHDLQL SEDEYNSERD SSLLAEFSDY GEISEDDEE DFMNAIREAS NFKVKKKKKN DKGKSYGRQR KERVLDPEVA QLLSQANEAF VRNDLQVAER LFNEVIKKDA R NFAAYETL ...String:
MAAGKLKKEQ QNQSAERESA DTGKVNDEDE EHLYGNIDDY KHLIQDEEYD DEDVPHDLQL SEDEYNSERD SSLLAEFSDY GEISEDDEE DFMNAIREAS NFKVKKKKKN DKGKSYGRQR KERVLDPEVA QLLSQANEAF VRNDLQVAER LFNEVIKKDA R NFAAYETL GDIYQLQGRL NDCCNSWFLA AHLNASDWEF WKIVAILSAD LDHVRQAIYC FSRVISLNPM EWESIYRRSM LY KKTGQLA RALDGFQRLY MYNPYDANIL RELAILYVDY DRIEDSIELY MKVFNANVER REAILAALEN ALDSSDEESA AEG EDADEK EPLEQDEDRQ MFPDINWKKI DAKYKCIPFD WSSLNILAEL FLKLAVSEVD GIKTIKKCAR WIQRRESQTF WDHV PDDSE FDNRRFKNST FDSLLAAEKE KSYNIPIDIR VRLGLLRLNT DNLVEALNHF QCLYDETFSD VADLYFEAAT ALTRA EKYK EAIDFFTPLL SLEEWRTTDV FKPLARCYKE IESYETAKEF YELAIKSEPD DLDIRVSLAE VYYRLNDPET FKHMLV DVV EMRKHQVDET LHRISNEKSS NDTSDISSKP LLEDSKFRTF RKKKRTPYDA ERERIERERR ITAKVVDKYE KMKKFEL NS GLNEAKQASI WINTVSELVD IFSSVKNFFM KSRSRKFVGI LRRTKKFNTE LDFQIERLSK LAEGDSVFEG PLMEERVT L TSATELRGLS YEQWFELFME LSLVIAKYQS VEDGLSVVET AQEVNVFFQD PERVKMMKFV KLAIVLQMDD EEELAENLR GLLNQFQFNR KVLQVFMYSL CRGPSSLNIL SSTIQQKFFL RQLKAFDSCR YNTEVNGQAS ITNKEVYNPN KKSSPYLYYI YAVLLYSSR GFLSALQYLT RLEEDIPDDP MVNLLMGLSH IHRAMQRLTA QRHFQIFHGL RYLYRYHKIR KSLYTDLEKQ E ADYNLGRA FHLIGLVSIA IEYYNRVLEN YDDGKLKKHA AYNSIIIYQQ SGNVELADHL MEKYLSI

UniProtKB: Transcription factor tau 131 kDa subunit

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Macromolecule #4: Transcription factor tau 95 kDa subunit

MacromoleculeName: Transcription factor tau 95 kDa subunit / type: protein_or_peptide / ID: 4 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 73.649406 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MPVEEPLATL SSIPDSSADQ APPLIADEFT LDLPRIPSLE LPLNVSTKHS SIQKAIKMCG GIEKVKEAFK EHGPIESQHG LQLYLNDDT DSDGSKSYFN EHPVIGKRVP FRDESVILKV TMPKGTLSKN NNSVKDSIKS LKDSNKLRVT PVSIVDNTIK F REMSDFQI ...String:
MPVEEPLATL SSIPDSSADQ APPLIADEFT LDLPRIPSLE LPLNVSTKHS SIQKAIKMCG GIEKVKEAFK EHGPIESQHG LQLYLNDDT DSDGSKSYFN EHPVIGKRVP FRDESVILKV TMPKGTLSKN NNSVKDSIKS LKDSNKLRVT PVSIVDNTIK F REMSDFQI KLDNVPSARE FKSSFGSLEW NNFKSFVNSV PDNDSQPQEN IGNLILDRSV KIPSTDFQLP PPPKLSMVGF PL LYKYKAN PFAKKKKNGV TEVKGTYIKN YQLFVHDLSD KTVIPSQAHE QVLYDFEVAK KTKVYPGTKS DSKFYESLEE CLK ILRELF ARRPIWVKRH LDGIVPKKIH HTMKIALALI SYRFTMGPWR NTYIKFGIDP RSSVEYAQYQ TEYFKIERKL LSSP IVKKN VPKPPPLVFE SDTPGGIDSR FKFDGKRIPW YLMLQIDLLI GEPNIAEVFH NVEYLDKANE LTGWFKELDL VKIRR IVKY ELGCMVQGNY EYNKYKLKYF KTMLFVKESM VPENKNSEEG MGVNTNKDAD GDINMDAGSQ MSSNAIEEDK GIAAGD DFD DNGAITEEPD DAALENEEMD TDQNLKVPAS IDDDVDDVDA DEEEQESFDV KTASFQDIIN KIAKLDPKTA ETMKSEL KG FVDEVDL

UniProtKB: Transcription factor tau 95 kDa subunit

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Macromolecule #5: Transcription factor tau 91 kDa subunit

MacromoleculeName: Transcription factor tau 91 kDa subunit / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 74.803297 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MAVIPAKKRG RPRKSVVAEV PYDSLASPVS ENSGSKRPRR NASKKAVANF AQLVHAGRDD VINTTQVNNV DDTDDDDFVL NDEGDGEES DNVEIEFENE LESTKNEVAD LNSSGSGASV RPSGRRNTVQ KLRLKKNSTK NMKSSSPGSS LGQKGRPIRL L KDLSSARD ...String:
MAVIPAKKRG RPRKSVVAEV PYDSLASPVS ENSGSKRPRR NASKKAVANF AQLVHAGRDD VINTTQVNNV DDTDDDDFVL NDEGDGEES DNVEIEFENE LESTKNEVAD LNSSGSGASV RPSGRRNTVQ KLRLKKNSTK NMKSSSPGSS LGQKGRPIRL L KDLSSARD KIERIYGLNK EKLLLLAKVK EGFETSVFDF PFKNIQPDSP YFVCLDPPCK KESAYNKVIG DKNRTVYHEI NK TEFENMI KLRTKRLKLL IGEVDAEVST GDKIEFPVLA NGKRRGFIYN VGGLVTDIAW LNIEENTDIG KDIQYLAVAV SQY MDEPLN EHLEMFDKEK HSSCIQIFKM NTSTLHCVKV QTIVHSFGEV WDLKWHEGCH APHLVGCLSF VSQEGTINFL EIID NATDV HVFKMCEKPS LTLSLADSLI TTFDFLSPTT VVCGFKNGFV AEFDLTDPEV PSFYDQVHDS YILSVSTAYS DFEDT VVST VAVDGYFYIF NPKDIATTKT TVSRFRGSNL VPVVYCPQIY SYIYSDGASS LRAVPSRAAF AVHPLVSRET TITAIG VSR LHPMVLAGSA DGSLIITNAA RRLLHGIKNS SATQKSLRLW KWDYSIKDDK YRIDSSYEVY PLTVNDVSKA KIDAHGI NI TCTKWNETSA GGKCYAFSNS AGLLTLEYLS

UniProtKB: Transcription factor tau 91 kDa subunit

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Macromolecule #6: Transcription factor tau 60 kDa subunit

MacromoleculeName: Transcription factor tau 60 kDa subunit / type: protein_or_peptide / ID: 6 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 67.755172 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MKLLKDLLVD RKEFEDWKNN LTWARDGTLY LTTFPDISIG QPKYAKDINC NSKNLFHVKE FPLEFENKLD FELAQQNGLL NSQPVCYPR VCKPSPIDDW MAVLSNNGNV SVFKDNKMLT NLDSKGNLSS RTYHCFEWNP IESSIVVGNE DGELQFFSIR K NSENTPEF ...String:
MKLLKDLLVD RKEFEDWKNN LTWARDGTLY LTTFPDISIG QPKYAKDINC NSKNLFHVKE FPLEFENKLD FELAQQNGLL NSQPVCYPR VCKPSPIDDW MAVLSNNGNV SVFKDNKMLT NLDSKGNLSS RTYHCFEWNP IESSIVVGNE DGELQFFSIR K NSENTPEF YFESSIRLSD AGSKDWVTHI VWYEDVLVAA LSNNSVFSMT VSASSHQPVS RMIQNASRRK ITDLKIVDYK VV LTCPGYV HKIDLKNYSI SSLKTGSLEN FHIIPLNHEK ESTILLMSNK TSYKVLLEDE LHVTADNIIA PYLEKKFKKW STI WNEFNN YETTLVIHGI SLSPDGYSIA IVYDMERVAF KYKIASEQSF NIMFAPLYHT WTISERAVGL AWYQTYQIYN QSLP KLPEN FSMNKKLLNG NYPISLDFQS YLNALMKSEE MRIIMFLNMT IDKPSILSFL EALYEYAINK KSELTNSFDL ACVLS IAAI LKREAPIYNG TLLMKNSFLE ETFNLESFTA DPETVTSTTN NTWKRCGVTL LPILTTHVKI CPVSKQRVID IKRDDL NDY GWFTRGLLER FNEISVYCGT TLEVM

UniProtKB: Transcription factor tau 60 kDa subunit

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Macromolecule #7: Transcription factor tau 55 kDa subunit

MacromoleculeName: Transcription factor tau 55 kDa subunit / type: protein_or_peptide / ID: 7 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 49.198898 KDa
Recombinant expressionOrganism: Saccharomyces cerevisiae (brewer's yeast)
SequenceString: MVVNTIYIAR HGYRSNWLPE GPYPDPLTGI DSDVPLAEHG VQQAKELAHY LLSLDNQPEA AFASPFYRCL ETVQPIAKLL EIPVYLERG IGEWYRPDRK PVIPVPAGYE ILSKFFPGVI SQEWDSTLTP NEKGETEQEM YMRFKKFWPL FIERVEKEYP N VECILLVT ...String:
MVVNTIYIAR HGYRSNWLPE GPYPDPLTGI DSDVPLAEHG VQQAKELAHY LLSLDNQPEA AFASPFYRCL ETVQPIAKLL EIPVYLERG IGEWYRPDRK PVIPVPAGYE ILSKFFPGVI SQEWDSTLTP NEKGETEQEM YMRFKKFWPL FIERVEKEYP N VECILLVT HAASKIALGM SLLGYDNPRM SLNENGDKIR SGSCSLDKYE ILKKSYDTID ETDDQTSFTY IPFSDRKWVL TM NGNTEFL SSGEEMNWNF DCVAEAGSDA DIKKRQMTKK TSSPIPEADD QTEVETVYIS VDIPSGNYKE RTEIAKSAIL QYS GLETDA PLFRIGNRLY EGSWERLVGT ELAFPNAAHV HKKTAGLLSP TEENETTNAG QSKGSSTAND PNIQIQEEDV GLPD STNTS RDHTGDKEEV QSEKIYRIKE RIVLSNVRPM

UniProtKB: Transcription factor tau 55 kDa subunit

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Macromolecule #8: Transcription factor IIIB 70 kDa subunit,TATA-box-binding protein

MacromoleculeName: Transcription factor IIIB 70 kDa subunit,TATA-box-binding protein
type: protein_or_peptide / ID: 8 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Saccharomyces cerevisiae S288C (yeast)
Molecular weightTheoretical: 82.097867 KDa
Recombinant expressionOrganism: Escherichia coli (E. coli)
SequenceString: MPVCKNCHGT EFERDLSNAN NDLVCKACGV VSEDNPIVSE VTFGETSAGA AVVQGSFIGA GQSHAAFGGS SALESREATL NNARRKLRA VSYALHIPEY ITDAAFQWYK LALANNFVQG RRSQNVIASC LYVACRKEKT HHMLIDFSSR LQVSVYSIGA T FLKMVKKL ...String:
MPVCKNCHGT EFERDLSNAN NDLVCKACGV VSEDNPIVSE VTFGETSAGA AVVQGSFIGA GQSHAAFGGS SALESREATL NNARRKLRA VSYALHIPEY ITDAAFQWYK LALANNFVQG RRSQNVIASC LYVACRKEKT HHMLIDFSSR LQVSVYSIGA T FLKMVKKL HITELPLADP SLFIQHFAEK LDLADKKIKV VKDAVKLAQR MSKDWMFEGR RPAGIAGACI LLACRMNNLR RT HTEIVAV SHVAEETLQQ RLNEFKNTKA AKLSVQKFRE NDVEDGEARP PSFVKNRKKE RKIKDSLDKE EMFQTSEEAL NKN PILTQV LGEQELSSKE VLFYLKQFSE RRARVVERIK ATNGIDGENI YHEGSENETR KRKLSEAMPW SGIVPTLQNI VATV TLGCR LDLKTVALHA RNAEYNPKRF AAVIMRIREP KTTALIFASG KMVVTGAKSE DDSKLASRKY ARIIQKIGFA AKFTD FKIQ NIVGSCDVKF PIRLEGLAFS HGTFSSYEPE LFPGLIYRMV KPKIVLLIFV SGKIVLTGAK QREEIYQAFE AIYPVL SEF RKMGSGSGSG SGSGSPRNLH LLPTTDTYLS KVSDDPDNLE DVDDEELNAH LLNEEASKLK ERIWIGLNAD FLLEQES KR LKQEADIATG NTSVKKKRTR RRNNTRSDEP TKTVDAAAAI GLMSDLQDKS GLHAALKAAE ESGDFTTADS VKNMLQKA S FSKKINYDAI DGLFR

UniProtKB: Transcription factor IIIB 70 kDa subunit, TATA-box-binding protein, Transcription factor IIIB 70 kDa subunit

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Macromolecule #9: DNA (151-MER)

MacromoleculeName: DNA (151-MER) / type: dna / ID: 9 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 52.636594 KDa
SequenceString: (DA)(DA)(DC)(DA)(DT)(DG)(DT)(DC)(DT)(DG) (DG)(DA)(DC)(DC)(DC)(DT)(DG)(DC)(DC)(DC) (DT)(DC)(DA)(DT)(DA)(DT)(DC)(DA)(DC) (DC)(DT)(DG)(DC)(DG)(DT)(DT)(DT)(DC)(DC) (DG) (DT)(DT)(DA)(DA)(DA)(DC) ...String:
(DA)(DA)(DC)(DA)(DT)(DG)(DT)(DC)(DT)(DG) (DG)(DA)(DC)(DC)(DC)(DT)(DG)(DC)(DC)(DC) (DT)(DC)(DA)(DT)(DA)(DT)(DC)(DA)(DC) (DC)(DT)(DG)(DC)(DG)(DT)(DT)(DT)(DC)(DC) (DG) (DT)(DT)(DA)(DA)(DA)(DC)(DT)(DA) (DT)(DC)(DG)(DG)(DT)(DT)(DG)(DC)(DG)(DG) (DC)(DC) (DA)(DT)(DA)(DT)(DC)(DT)(DA) (DC)(DC)(DA)(DG)(DA)(DA)(DA)(DG)(DC)(DA) (DC)(DC)(DG) (DT)(DT)(DT)(DC)(DC)(DC) (DG)(DT)(DC)(DC)(DG)(DA)(DT)(DC)(DA)(DA) (DC)(DT)(DG)(DT) (DA)(DG)(DT)(DT)(DA) (DA)(DG)(DC)(DT)(DG)(DG)(DT)(DA)(DA)(DG) (DA)(DG)(DC)(DC)(DT) (DG)(DA)(DC)(DC) (DG)(DA)(DG)(DT)(DA)(DG)(DT)(DG)(DT)(DA) (DG)(DT)(DG)(DG)(DG)(DT) (DG)(DA)(DC) (DC)(DA)(DT)(DA)(DC)(DG)(DC)(DG)(DA)(DA) (DA)(DC)(DT)(DC)(DA)(DG)(DG) (DT)(DG) (DC)(DT)(DG)(DC)(DA)(DA)(DT)(DC)(DT)

GENBANK: GENBANK: CP011821.1

+
Macromolecule #10: DNA (151-MER)

MacromoleculeName: DNA (151-MER) / type: dna / ID: 10 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Saccharomyces cerevisiae (brewer's yeast)
Molecular weightTheoretical: 52.934797 KDa
SequenceString: (DA)(DG)(DA)(DT)(DT)(DG)(DC)(DA)(DG)(DC) (DA)(DC)(DC)(DT)(DG)(DA)(DG)(DT)(DT)(DT) (DC)(DG)(DC)(DG)(DT)(DA)(DT)(DG)(DG) (DT)(DC)(DA)(DC)(DC)(DC)(DA)(DC)(DT)(DA) (DC) (DA)(DC)(DT)(DA)(DC)(DT) ...String:
(DA)(DG)(DA)(DT)(DT)(DG)(DC)(DA)(DG)(DC) (DA)(DC)(DC)(DT)(DG)(DA)(DG)(DT)(DT)(DT) (DC)(DG)(DC)(DG)(DT)(DA)(DT)(DG)(DG) (DT)(DC)(DA)(DC)(DC)(DC)(DA)(DC)(DT)(DA) (DC) (DA)(DC)(DT)(DA)(DC)(DT)(DC)(DG) (DG)(DT)(DC)(DA)(DG)(DG)(DC)(DT)(DC)(DT) (DT)(DA) (DC)(DC)(DA)(DG)(DC)(DT)(DT) (DA)(DA)(DC)(DT)(DA)(DC)(DA)(DG)(DT)(DT) (DG)(DA)(DT) (DC)(DG)(DG)(DA)(DC)(DG) (DG)(DG)(DA)(DA)(DA)(DC)(DG)(DG)(DT)(DG) (DC)(DT)(DT)(DT) (DC)(DT)(DG)(DG)(DT) (DA)(DG)(DA)(DT)(DA)(DT)(DG)(DG)(DC)(DC) (DG)(DC)(DA)(DA)(DC) (DC)(DG)(DA)(DT) (DA)(DG)(DT)(DT)(DT)(DA)(DA)(DC)(DG)(DG) (DA)(DA)(DA)(DC)(DG)(DC) (DA)(DG)(DG) (DT)(DG)(DA)(DT)(DA)(DT)(DG)(DA)(DG)(DG) (DG)(DC)(DA)(DG)(DG)(DG)(DT) (DC)(DC) (DA)(DG)(DA)(DC)(DA)(DT)(DG)(DT)(DT)

GENBANK: GENBANK: CP011821.1

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Macromolecule #11: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 11 / Number of copies: 9 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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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

Concentration0.02 mg/mL
BufferpH: 7.9
GridModel: Quantifoil R2/1 / Material: COPPER / Mesh: 300 / Support film - Material: GRAPHENE OXIDE / Support film - topology: CONTINUOUS / Pretreatment - Type: GLOW DISCHARGE / Pretreatment - Time: 10 sec. / Pretreatment - Atmosphere: AIR
VitrificationCryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 277.15 K / Instrument: FEI VITROBOT MARK IV

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Image recordingFilm or detector model: GATAN K3 (6k x 4k) / Average electron dose: 60.0 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 5.0 µm / Nominal defocus min: 2.0 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

+
Image processing

Particle selectionNumber selected: 5748589
Startup modelType of model: OTHER / Details: Map from negatively stained sample
Final reconstructionNumber classes used: 1 / Resolution.type: BY AUTHOR / Resolution: 3.8 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: RELION (ver. 3.1) / Number images used: 78512
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: RELION (ver. 3.1)
Final 3D classificationNumber classes: 3 / Avg.num./class: 75000 / Software - Name: RELION (ver. 3.1)
FSC plot (resolution estimation)

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Atomic model buiding 1

RefinementSpace: REAL / Protocol: RIGID BODY FIT
Output model

PDB-8ffz:
TFIIIA-TFIIIC-Brf1-TBP complex bound to 5S rRNA gene

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