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- EMDB-51275: Cryo-EM structure of Vibrio cholerae RNA polymerase Transcription... -

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Basic information

Entry
Database: EMDB / ID: EMD-51275
TitleCryo-EM structure of Vibrio cholerae RNA polymerase Transcription Activation Complex with ToxR transcription factor and ompU promoter DNA
Map dataTAC_ompU MAP D. Composite map.
Sample
  • Complex: Virulent transcription complex #3
    • Complex: DNA-directed RNA polymerase
      • Protein or peptide: DNA-directed RNA polymerase subunit beta
      • Protein or peptide: DNA-directed RNA polymerase subunit beta'
      • Protein or peptide: DNA-directed RNA polymerase subunit omega
      • Protein or peptide: DNA-directed RNA polymerase subunit alpha
      • Protein or peptide: RNA polymerase sigma factor RpoD
      • Protein or peptide: Cholera toxin transcriptional activator
    • Complex: Double stranded DNA
      • DNA: ompU promoter template DNA strand
      • DNA: ompU promoter non-template DNA strand
  • Ligand: ZINC ION
  • Ligand: MAGNESIUM ION
KeywordsRNA polymerase / Open promoter complex / ompU / Vibrio cholerae / TRANSCRIPTION
Function / homology
Function and homology information


phosphorelay response regulator activity / sigma factor activity / DNA-directed RNA polymerase complex / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / protein dimerization activity / DNA-templated transcription / regulation of DNA-templated transcription ...phosphorelay response regulator activity / sigma factor activity / DNA-directed RNA polymerase complex / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / protein dimerization activity / DNA-templated transcription / regulation of DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / plasma membrane / cytoplasm
Similarity search - Function
OmpR/PhoB-type DNA-binding domain / Transcriptional regulatory protein, C terminal / OmpR/PhoB-type DNA-binding domain profile. / Transcriptional regulatory protein, C terminal / RNA polymerase sigma factor 70, non-essential domain / Sigma-70, non-essential region / RNA polymerase sigma factor 70, region 1.1 / Sigma-70 factor, region 1.1 superfamily / Sigma-70 factor, region 1.1 / Signal transduction response regulator, C-terminal effector ...OmpR/PhoB-type DNA-binding domain / Transcriptional regulatory protein, C terminal / OmpR/PhoB-type DNA-binding domain profile. / Transcriptional regulatory protein, C terminal / RNA polymerase sigma factor 70, non-essential domain / Sigma-70, non-essential region / RNA polymerase sigma factor 70, region 1.1 / Sigma-70 factor, region 1.1 superfamily / Sigma-70 factor, region 1.1 / Signal transduction response regulator, C-terminal effector / Sigma-70 factors family signature 1. / RNA polymerase sigma factor RpoD, C-terminal / RNA polymerase sigma factor RpoD / : / RNA polymerase sigma-70 region 1.2 / Sigma-70 factor, region 1.2 / RNA polymerase sigma-70 region 3 / Sigma-70 region 3 / Sigma-70 factors family signature 2. / RNA polymerase sigma-70 / RNA polymerase sigma-70 region 4 / Sigma-70, region 4 / RNA polymerase sigma-70 region 2 / RNA polymerase sigma-70 like domain / Sigma-70 region 2 / RNA polymerase sigma factor, region 2 / RNA polymerase sigma factor, region 3/4-like / DNA-directed RNA polymerase, omega subunit / DNA-directed RNA polymerase, subunit beta-prime, bacterial type / DNA-directed RNA polymerase, beta subunit, external 1 domain superfamily / DNA-directed RNA polymerase, beta subunit, external 1 domain / RNA polymerase beta subunit external 1 domain / RNA polymerase, alpha subunit, C-terminal / Bacterial RNA polymerase, alpha chain C terminal domain / DNA-directed RNA polymerase beta subunit, bacterial-type / DNA-directed RNA polymerase, alpha subunit / DNA-directed RNA polymerase, subunit beta-prime / RNA polymerase Rpb6 / RNA polymerase, subunit omega/Rpo6/RPB6 / RNA polymerase Rpb6 / RNA polymerase Rpb1, domain 3 superfamily / RPB6/omega subunit-like superfamily / DNA-directed RNA polymerase, insert domain / DNA-directed RNA polymerase, RpoA/D/Rpb3-type / RNA polymerase Rpb3/RpoA insert domain / RNA polymerase Rpb3/Rpb11 dimerisation domain / RNA polymerases D / RNA polymerase Rpb1, clamp domain superfamily / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, domain 1 / RNA polymerase, beta subunit, protrusion / RNA polymerase beta subunit / RNA polymerase Rpb1, domain 1 / RNA polymerase, alpha subunit / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 4 / RNA polymerase Rpb1, domain 2 / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 4 / DNA-directed RNA polymerase, insert domain superfamily / RNA polymerase Rpb2, domain 2 / RNA polymerase Rpb2, domain 2 / RNA polymerase, N-terminal / RNA polymerase Rpb1, funnel domain superfamily / RNA polymerase I subunit A N-terminus / RNA polymerase, RBP11-like subunit / RNA polymerase, beta subunit, conserved site / RNA polymerase Rpb2, domain 7 / RNA polymerase Rpb2, domain 3 / RNA polymerase Rpb2, OB-fold / RNA polymerase Rpb2, domain 7 / RNA polymerase Rpb2, domain 3 / RNA polymerases beta chain signature. / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain / DNA-directed RNA polymerase, subunit 2 / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain superfamily / RNA polymerase Rpb2, domain 6 / Winged helix-like DNA-binding domain superfamily
Similarity search - Domain/homology
RNA polymerase sigma factor RpoD / DNA-directed RNA polymerase subunit beta / DNA-directed RNA polymerase subunit beta' / DNA-directed RNA polymerase subunit omega / DNA-directed RNA polymerase subunit alpha / Cholera toxin transcriptional activator
Similarity search - Component
Biological speciesVibrio cholerae O395 (bacteria) / Vibrio cholerae O1 biovar El Tor str. N16961 (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 3.1 Å
AuthorsAlcaide-Jimenez A / Baudin F / Canals A / Machon C / Murciano B / Fabrega-Ferrer M / Bantysh O / Perez-Luque R / Krukonis ES / Muller CW / Coll M
Funding support Spain, 2 items
OrganizationGrant numberCountry
Ministerio de Ciencia e Innovacion (MCIN)PID2020-12141GB-100 Spain
Generalitat de Catalunya2021SGR00423 Spain
CitationJournal: Sci Adv / Year: 2026
Title: Structures of transcription complexes reveal how ToxR and TcpP recruit the RNA polymerase and activate virulence genes.
Authors: Adrià Alcaide-Jiménez / Albert Canals / Florence Baudin / Cristina Machón / Montserrat Fàbrega-Ferrer / Olga Bantysh / Rosa Pérez-Luque / Brice Murciano / Ali A Mohammad / Michael J ...Authors: Adrià Alcaide-Jiménez / Albert Canals / Florence Baudin / Cristina Machón / Montserrat Fàbrega-Ferrer / Olga Bantysh / Rosa Pérez-Luque / Brice Murciano / Ali A Mohammad / Michael J Rowse / Joseph M Ferracciolo / Eric S Krukonis / Christoph W Müller / Miquel Coll /
Abstract: Activation of virulence in , the etiological agent of cholera disease, is mediated by two transmembrane one-component signal-transduction proteins, ToxR and TcpP, which are also transcription factors. ...Activation of virulence in , the etiological agent of cholera disease, is mediated by two transmembrane one-component signal-transduction proteins, ToxR and TcpP, which are also transcription factors. Using cryo-electron microscopy, we have solved five structures of the and transcription activation complexes, including the RNA polymerase (RNAP) holoenzyme, promoter DNAs, transcribed RNA, and their corresponding transcription factors, ToxR or TcpP and ToxR-TcpP, respectively. Activation is achieved through the interaction of ToxR or TcpP with the α-C-terminal repeat domain of RNAP where a single residue of the activator, a phenylalanine, appears to be the most critical contact, as confirmed by mutagenesis. No interactions of the transcription factors were observed with other subunits of the RNAP, i.e., the σ subunit as it occurs in the structurally related PhoB family of two-component transcription factors. The structures, and their comparison with our previously solved DNA promoter-ToxR x-ray structures, unveil the molecular mechanism of cholera virulence gene activation.
History
DepositionAug 6, 2024-
Header (metadata) releaseDec 17, 2025-
Map releaseDec 17, 2025-
UpdateJan 28, 2026-
Current statusJan 28, 2026Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_51275.png

Downloads & links

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Map

FileDownload / File: emd_51275.map.gz / Format: CCP4 / Size: 347.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationTAC_ompU MAP D. Composite map.
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
0.82 Å/pix.
x 450 pix.
= 369.9 Å		0.82 Å/pix.
x 450 pix.
= 369.9 Å		0.82 Å/pix.
x 450 pix.
= 369.9 Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 0.822 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.0996
Minimum - Maximum-0.013458796 - 2.0014844
Average (Standard dev.)0.0016090198 (±0.025426243)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions450450450
Spacing450450450
CellA=B=C: 369.90002 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Virulent transcription complex #3

EntireName: Virulent transcription complex #3
Components
  • Complex: Virulent transcription complex #3
    • Complex: DNA-directed RNA polymerase
      • Protein or peptide: DNA-directed RNA polymerase subunit beta
      • Protein or peptide: DNA-directed RNA polymerase subunit beta'
      • Protein or peptide: DNA-directed RNA polymerase subunit omega
      • Protein or peptide: DNA-directed RNA polymerase subunit alpha
      • Protein or peptide: RNA polymerase sigma factor RpoD
      • Protein or peptide: Cholera toxin transcriptional activator
    • Complex: Double stranded DNA
      • DNA: ompU promoter template DNA strand
      • DNA: ompU promoter non-template DNA strand
  • Ligand: ZINC ION
  • Ligand: MAGNESIUM ION

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Supramolecule #1: Virulent transcription complex #3

SupramoleculeName: Virulent transcription complex #3 / type: complex / ID: 1 / Parent: 0 / Macromolecule list: #1-#8

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Supramolecule #2: DNA-directed RNA polymerase

SupramoleculeName: DNA-directed RNA polymerase / type: complex / ID: 2 / Parent: 1 / Macromolecule list: #1-#6
Source (natural)Organism: Vibrio cholerae O395 (bacteria)

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Supramolecule #3: Double stranded DNA

SupramoleculeName: Double stranded DNA / type: complex / ID: 3 / Parent: 1 / Macromolecule list: #7-#8
Source (natural)Organism: Vibrio cholerae O395 (bacteria)

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Macromolecule #1: DNA-directed RNA polymerase subunit beta

MacromoleculeName: DNA-directed RNA polymerase subunit beta / type: protein_or_peptide / ID: 1 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 149.696531 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString: MVYSYTEKKR IRKDFGTRPQ VLDIPYLLSI QLDSFEKFIE QDPEGQYGLE AAFRSVFPIQ SYNGNSELQY VSYRLGEPVF DVKECQIRG VTYSKPLRVK LRLVIFDKDA PAGTVKDIKE QEVYMGEIPL MTENGTFVIN GTERVIVSQL HRSPGVFFDS D KGKTHSSG ...String:
MVYSYTEKKR IRKDFGTRPQ VLDIPYLLSI QLDSFEKFIE QDPEGQYGLE AAFRSVFPIQ SYNGNSELQY VSYRLGEPVF DVKECQIRG VTYSKPLRVK LRLVIFDKDA PAGTVKDIKE QEVYMGEIPL MTENGTFVIN GTERVIVSQL HRSPGVFFDS D KGKTHSSG KVLYNARIIP YRGSWLDFEF DPKDNLYVRI DRRRKLPASI ILRVLGKTSA EILDIFFEKV NFEVKDQTLM ME LVPERLR GETATFDIEA DGKVYVEKGR RVTARHIRQL EKDGVNFIEV PVEYIVGKVS AKDYVNEATG ELIITANQEI SLE ALANLS QAGYKKLEVL FTNDLDHGPF MSETLRVDST TDRISALVEI YRMMRPGEPP TKEAAESLFE SLFFSAERYD LSTV GRMKF NSSIGREDAE EQGTLDEVDI IEVMKKLISI RNGKGEVDDI DHLGNRRIRS VGEMAENQFR VGLVRVERAV KERLS LGDL DNVMPQDLIN AKPISAAVKE FFGSSQLSQF MDQNNPLSEV THKRRISALG PGGLTRERAG FEVRDVHVTH YGRLCP IET PEGPNIGLIN SLSAFARCNE YGFLETPYRR VVNGVVTDEV DYLSAIEEGQ FVIAQANAKL TEEGGFADEL VTARQKG ES GLHPREHVDY MDVATNQVVS IAASLIPFLE HDDANRALMG ANMQRQAVPT LRSEKPLVGT GIERNVAVDS GVTAVAKR G GVIQSVDASR IVVKVNEEEL IPGEAGIDIY NLTKYTRSNQ NTCINQRPCV MPGEPVARGD VLADGPSTDL GELALGQNM RIAFMPWNGY NFEDSILVSE RVVQDDRFTT IHIQELSCVA RDTKLGAEEI TADIPNVGEA ALSKLDESGI VYIGAEVKGG DILVGKVTP KGETQLTPEE KLLRAIFGEK ASDVKDTSLR VPNSVAGTVI DVQVFTRDGV EKDKRALEIE QMQLKEAKKD L TEEFQILE GGLLARVRSV LLAGGYTEAK LSSIERKKWL EQTLENEELQ NQLEQLAEQY DELKADFDKK FEAKRRKITQ GD DLAPGVL KIVKVYLAVK RRIQPGDKMA GRHGNKGVIS KINPVEDMPY DENGQPVDIV LNPLGVPSRM NIGQILEVHL GLA AKGIGD KINQMIKEQQ ELAKLREFLQ KVYDLGDTRQ RVDISELSDE DVRTLAHNLR AGLPVATPVF DGAPESSIKA MLEL ADLPA SGQLTLFDGR TGDAFERPVT VGYMYMLKLN HLVDDKMHAR STGSYSLVTQ QPLGGKAQFG GQRFGEMEVW ALEAY GAAY TLQEMLTVKS DDVNGRTKMY KNIVDGNHAM EPGMPESFNV LLKEIRSLGI NIELEDE

UniProtKB: DNA-directed RNA polymerase subunit beta

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Macromolecule #2: DNA-directed RNA polymerase subunit beta'

MacromoleculeName: DNA-directed RNA polymerase subunit beta' / type: protein_or_peptide / ID: 2 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 155.234281 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString: MKDLLNFLKA QHKTEEFDAI KIGLASPDMI RSWSFGEVKK PETINYRTFK PERDGLFCAR IFGPVKDYEC LCGKYKRLKH RGVICEKCG VEVTQTKVRR DRMGHIELAS PVAHIWFLKS LPSRIGLLMD MPLRDIERVL YFEMYVVTEP GMTDLERGQM L TEEEYLDR ...String:
MKDLLNFLKA QHKTEEFDAI KIGLASPDMI RSWSFGEVKK PETINYRTFK PERDGLFCAR IFGPVKDYEC LCGKYKRLKH RGVICEKCG VEVTQTKVRR DRMGHIELAS PVAHIWFLKS LPSRIGLLMD MPLRDIERVL YFEMYVVTEP GMTDLERGQM L TEEEYLDR LEEWGDEFTA KMGAEAIKDL LASMDLPAEA EQMREELDTT NSETKRKKLT KRLKLVEAFV ASGNKPEWMI LT VLPVLPP DLRPLVPLDG GRFATSDLND LYRRVINRNN RLKRLLELAA PDIIVRNEKR MLQESVDALL DNGRRGRAIT GSN KRPLKS LADMIKGKQG RFRQNLLGKR VDYSGRSVIT VGPYLRLHQC GLPKKMALEL FKPFIYSKLE TRGLATTIKA AKKM VEREE AVVWDILDEV IREHPVLLNR APTLHRLGIQ AFEPVLIEGK AIQLHPLVCA AYNADFDGDQ MAVHVPLTLE AQLEA RTLM MSTNNILSPA SGDPIIVPSQ DVVLGLYYMT REKINAKGEG MYLTGPAEAE KAYRTKTAEL HARVKVRITE TIKHEN GKL TTETKMIDTT VGRAMLWQIV PKGLPYSLVN QKLGKKQISN LLNEAYRKLG LKDTVIFADQ IMYTGFAYAA LSGVSVG ID DMVVPAAKYT EIAEAEEEVR EIQEQFQSGL VTAGERYNKV IDIWASTNDR VAKAMMENLS SEQVINRQGE QEKQESFN S IYMMADSGAR GSAAQIRQLA GMRGLMARPD GSIIETPITA NFKEGLNVLQ YFISTHGARK GLADTALKTA NSGYLTRRL VDVAQDVVVT EHDCGTLEGV VMTPHIEGGD VKVALTELAL GRVVSEDILK PGTDEVLIPR NTLLDEKWCK VINDNSVDQI KVRSVVTCD SDFGCCAQCY GRDLARGHLV NQGEAVGVIA AQSIGEPGTQ LTMRTFHIGG AASTAAAENS IQAKNNGSVK L HNAKFVTN KDGKLVITSR ASELTIIDEF GRTKEKHKLP YGSMLSKADG DAVAAGETVA NWEAHTMPII TEVAGRVQFV DM IDGVTVS RQTDDLTGLS SSEVTEAAAR PAAGKDMRPA IKLVDANGKD VLIPGTDMPA QYFLPGKAIV NLDDGAEVNV GDT LARIPQ KSGGNKDITG GLPRVADLFE ARKPKEPAIL AEHSGTVSFG KETKGKRRLI ITRDSGDTYE EMIPKHRQLN VFEG ERIER GDVIADGPES PHDILRLRGI HAVTTYIANE VQEVYRLQGV KINDKHIETI VRQMLRKCTI TFAGDSEFLP GETVE YSQV KIANRKLVEE GKEPARFERE LLGITKASLA TESFISAASF QETTRVLTEA AVSGKRDDLR GLKENVIVGR LIPAGT GFA YHQDRQAKRA QEQQGPSAEQ ATDNLAALLN AGFSSDDE

UniProtKB: DNA-directed RNA polymerase subunit beta'

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Macromolecule #3: DNA-directed RNA polymerase subunit omega

MacromoleculeName: DNA-directed RNA polymerase subunit omega / type: protein_or_peptide / ID: 3 / Number of copies: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 10.068355 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString:
MARVTVQDAV EKIGNRFDLV LVAARRARQM QSGGKDALVP EENDKPTVIA LREIEEGLIT KDVLDARERQ EQQEQEAAEL AAVSSIMHN R

UniProtKB: DNA-directed RNA polymerase subunit omega

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Macromolecule #4: DNA-directed RNA polymerase subunit alpha

MacromoleculeName: DNA-directed RNA polymerase subunit alpha / type: protein_or_peptide / ID: 4 / Number of copies: 3 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 36.461473 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString: MQGSVTEFLK PRLVDIEQIS TTHAKVTLEP LERGFGHTLG NALRRILLSS MPGCAVTEVE IEGVLHEYST KEGVQEDILE ILLNLKGLA VRVAEGKDEV FITLNKSGSG PVVAGDITHD GDVEIVNPEH VICHLTSDNA AIAMRIKVER GRGYVPASAR I HTEEDERP ...String:
MQGSVTEFLK PRLVDIEQIS TTHAKVTLEP LERGFGHTLG NALRRILLSS MPGCAVTEVE IEGVLHEYST KEGVQEDILE ILLNLKGLA VRVAEGKDEV FITLNKSGSG PVVAGDITHD GDVEIVNPEH VICHLTSDNA AIAMRIKVER GRGYVPASAR I HTEEDERP IGRLLVDATF SPVDKIAYSV EAARVEQRTD LDKLVIDMET NGTLEPEEAI RRAATILAEQ LDAFVDLRDV RV PEEKEEK PEFDPILLRP VDDLELTVRS ANCLKAEAIH YIGDLVQRTE VELLKTPNLG KKSLTEIKDV LASRGLSLGM RLE NWPPAS IAED

UniProtKB: DNA-directed RNA polymerase subunit alpha

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Macromolecule #5: RNA polymerase sigma factor RpoD

MacromoleculeName: RNA polymerase sigma factor RpoD / type: protein_or_peptide / ID: 5 / Number of copies: 1 / Enantiomer: LEVO
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 70.749648 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString: MDQNPQSQLK QLVLRGKEQG YLTYAEVNDH LPAEIVDSEQ VEDIIQMIND MGIKVVETAP DADDLALSDD TTITDEDAAE AAAAALSSV ESEIGRTTDP VRMYMREMGT VELLTREGEI DIAKRIEDGI NQVQSAIAEY PGTIPYILEQ FDRVQAEELR L TDLISGFV ...String:
MDQNPQSQLK QLVLRGKEQG YLTYAEVNDH LPAEIVDSEQ VEDIIQMIND MGIKVVETAP DADDLALSDD TTITDEDAAE AAAAALSSV ESEIGRTTDP VRMYMREMGT VELLTREGEI DIAKRIEDGI NQVQSAIAEY PGTIPYILEQ FDRVQAEELR L TDLISGFV DPNDMETEAP TATHIGSELS EADLADEDDA VVEDEDEDGD GESSDSEEEV GIDPELAREK FNELRGKFQN LQ LAVNEFG RDSHQASEAS DLVLDIFREF RLTPKQFDHL VETLRTSMDR VRTQERLVMK AVVEVAKMPK KSFIALFTGN ESN EEWLDK VLASDKPYVA KVREQEEEIR RSIQKLQMIE QETSLSVERI KDISHRMSIG EAKARRAKKE MVEANLRLVI SIAK KYTNR GLQFLDLIQE GNIGLMKAVD KFEYRRGYKF STYATWWIRQ AITRSIADQA RTIRIPVHMI ETINKLNRIS RQMLQ EMGR EPLPEELAER MQMPEDKIRK VLKIAKEPIS METPIGDDED SHLGDFIEDT TLELPLDSAT ATSLKAATRD VLAGLT PRE AKVLRMRFGI DMNTDHTLEE VGKQFDVTRE RIRQIEAKAL RKLRHPSRSE VLRSFLDE

UniProtKB: RNA polymerase sigma factor RpoD

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Macromolecule #6: Cholera toxin transcriptional activator

MacromoleculeName: Cholera toxin transcriptional activator / type: protein_or_peptide / ID: 6 / Number of copies: 2 / Enantiomer: LEVO
Source (natural)Organism: Vibrio cholerae O1 biovar El Tor str. N16961 (bacteria)
Molecular weightTheoretical: 12.832575 KDa
Recombinant expressionOrganism: Escherichia coli BL21(DE3) (bacteria)
SequenceString:
MKFILAEKFT FDPLSNTLID KEDSEEIIRL GSNESRILWL LAQRPNEVIS RNDLHDFVWR EQGFEVDDSS LTQAISTLRK MLKDSTKSP QYVKTVPKRG YQLIARVETV E

UniProtKB: Cholera toxin transcriptional activator

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Macromolecule #7: ompU promoter template DNA strand

MacromoleculeName: ompU promoter template DNA strand / type: dna / ID: 7 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 21.146613 KDa
SequenceString: (DG)(DC)(DC)(DA)(DT)(DT)(DT)(DT)(DC)(DG) (DG)(DC)(DC)(DT)(DC)(DT)(DG)(DT)(DG)(DA) (DA)(DA)(DA)(DA)(DC)(DT)(DC)(DC)(DC) (DA)(DA)(DA)(DG)(DT)(DT)(DC)(DC)(DG)(DC) (DT) (DA)(DT)(DC)(DA)(DG)(DT) ...String:
(DG)(DC)(DC)(DA)(DT)(DT)(DT)(DT)(DC)(DG) (DG)(DC)(DC)(DT)(DC)(DT)(DG)(DT)(DG)(DA) (DA)(DA)(DA)(DA)(DC)(DT)(DC)(DC)(DC) (DA)(DA)(DA)(DG)(DT)(DT)(DC)(DC)(DG)(DC) (DT) (DA)(DT)(DC)(DA)(DG)(DT)(DT)(DA) (DG)(DT)(DA)(DA)(DA)(DA)(DT)(DG)(DA)(DT) (DA)(DT) (DA)(DA)(DA)(DT)(DC)(DC)(DA) (DA)(DC)

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Macromolecule #8: ompU promoter non-template DNA strand

MacromoleculeName: ompU promoter non-template DNA strand / type: dna / ID: 8 / Number of copies: 1 / Classification: DNA
Source (natural)Organism: Vibrio cholerae O395 (bacteria)
Molecular weightTheoretical: 21.408746 KDa
SequenceString: (DG)(DT)(DT)(DG)(DG)(DA)(DT)(DT)(DT)(DA) (DT)(DA)(DT)(DC)(DA)(DT)(DT)(DT)(DT)(DA) (DC)(DT)(DA)(DA)(DC)(DT)(DG)(DA)(DT) (DA)(DG)(DC)(DG)(DG)(DA)(DA)(DC)(DT)(DT) (DT) (DG)(DG)(DG)(DA)(DG)(DT) ...String:
(DG)(DT)(DT)(DG)(DG)(DA)(DT)(DT)(DT)(DA) (DT)(DA)(DT)(DC)(DA)(DT)(DT)(DT)(DT)(DA) (DC)(DT)(DA)(DA)(DC)(DT)(DG)(DA)(DT) (DA)(DG)(DC)(DG)(DG)(DA)(DA)(DC)(DT)(DT) (DT) (DG)(DG)(DG)(DA)(DG)(DT)(DA)(DA) (DA)(DA)(DG)(DA)(DG)(DT)(DC)(DT)(DC)(DC) (DG)(DC) (DG)(DA)(DA)(DA)(DA)(DT)(DG) (DG)(DC)

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Macromolecule #9: ZINC ION

MacromoleculeName: ZINC ION / type: ligand / ID: 9 / Number of copies: 2 / Formula: ZN
Molecular weightTheoretical: 65.409 Da

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Macromolecule #10: MAGNESIUM ION

MacromoleculeName: MAGNESIUM ION / type: ligand / ID: 10 / Number of copies: 1 / Formula: MG
Molecular weightTheoretical: 24.305 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

Concentration0.4 mg/mL
BufferpH: 8
Details: 10 mM HEPES pH 8.0, 500 mM NaCl, 1 mM DTT and 0.1 mM EDTA
VitrificationCryogen name: ETHANE

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Electron microscopy

MicroscopeTFS KRIOS
Image recordingFilm or detector model: GATAN K3 BIOQUANTUM (6k x 4k) / Average electron dose: 51.24 e/Å2
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 0.7000000000000001 µm
Sample stageSpecimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
Startup modelType of model: NONE
Final reconstructionResolution.type: BY AUTHOR / Resolution: 3.1 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 158389
Initial angle assignmentType: MAXIMUM LIKELIHOOD
Final angle assignmentType: MAXIMUM LIKELIHOOD
FSC plot (resolution estimation)

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