[English] 日本語
Yorodumi
- EMDB-14378: Cryo-EM structure of Mycobacterium tuberculosis RNA polymerase ho... -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: EMDB / ID: EMD-14378
TitleCryo-EM structure of Mycobacterium tuberculosis RNA polymerase holoenzyme dimer comprising sigma factor SigB
Map dataRNA polymerase dimer Class 1 (conformation 1)
Sample
  • Complex: RNA polymerase holoenzyme with sigma factor sigB
    • Protein or peptide: DNA-directed RNA polymerase subunit alphaPolymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit alphaPolymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit betaPolymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit beta'Polymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit omegaPolymerase
    • Protein or peptide: RNA polymerase sigma factor SigB
Biological speciesMycobacterium tuberculosis H37Rv (bacteria)
Methodsingle particle reconstruction / cryo EM / Resolution: 4.38 Å
AuthorsBrodolin K
Funding support France, 1 items
OrganizationGrant numberCountry
Agence Nationale de la Recherche (ANR)ANR-16-CE11-0025 France
Citation
Journal: Nucleic Acids Res / Year: 2014
Title: Mycobacterium RbpA cooperates with the stress-response σB subunit of RNA polymerase in promoter DNA unwinding.
Authors: Yangbo Hu / Zakia Morichaud / Ayyappasamy Sudalaiyadum Perumal / Françoise Roquet-Baneres / Konstantin Brodolin /
Abstract: RbpA, a transcriptional activator that is essential for Mycobacterium tuberculosis replication and survival during antibiotic treatment, binds to RNA polymerase (RNAP) in the absence of promoter DNA. ...RbpA, a transcriptional activator that is essential for Mycobacterium tuberculosis replication and survival during antibiotic treatment, binds to RNA polymerase (RNAP) in the absence of promoter DNA. It has been hypothesized that RbpA stimulates housekeeping gene expression by promoting assembly of the σ(A) subunit with core RNAP. Here, using a purified in vitro transcription system of M. tuberculosis, we show that RbpA functions in a promoter-dependent manner as a companion of RNAP essential for promoter DNA unwinding and formation of the catalytically active open promoter complex (RPo). Screening for RbpA activity using a full panel of the M. tuberculosis σ subunits demonstrated that RbpA targets σ(A) and stress-response σ(B), but not the alternative σ subunits from the groups 3 and 4. In contrast to σ(A), the σ(B) subunit activity displayed stringent dependency upon RbpA. These results suggest that RbpA-dependent control of RPo formation provides a mechanism for tuning gene expression during the switch between different physiological states, and in the stress response.
#1: Journal: Biorxiv / Year: 2022
Title: Structural basis of the mycobacterial stress-response RNA polymerase auto-inhibition via oligomerization
Authors: Morichaud Z / Trapani S / Vishwakarma R / Chaloin L / Lionne C / Lai-Kee-Him J / Bron P / Brodolin K
History
DepositionFeb 18, 2022-
Header (metadata) releaseFeb 8, 2023-
Map releaseFeb 8, 2023-
UpdateFeb 8, 2023-
Current statusFeb 8, 2023Processing site: PDBe / Status: Released

-
Structure visualization

Supplemental images

emd_14378.png

Downloads & links

-
Map

FileDownload / File: emd_14378.map.gz / Format: CCP4 / Size: 347.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
AnnotationRNA polymerase dimer Class 1 (conformation 1)
Projections & slices

Image control

Size
Brightness
Contrast
Others
AxesZ (Sec.)Y (Row.)X (Col.)
1.1 Å/pix.
x 450 pix.
= 495. Å		1.1 Å/pix.
x 450 pix.
= 495. Å		1.1 Å/pix.
x 450 pix.
= 495. Å

Surface

Projections

Slices (1/3)

Slices (1/2)

Slices (2/3)

Images are generated by Spider.

Voxel sizeX=Y=Z: 1.1 Å
Density

Histogram

Histogram (log scale)
Contour LevelBy AUTHOR: 0.25
Minimum - Maximum-0.3361331 - 1.2366912
Average (Standard dev.)-0.0018320102 (±0.04113083)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin000
Dimensions450450450
Spacing450450450
CellA=B=C: 495.0 Å
α=β=γ: 90.0 °

-
Supplemental data

-
Sample components

-
Entire : RNA polymerase holoenzyme with sigma factor sigB

EntireName: RNA polymerase holoenzyme with sigma factor sigB
Components
  • Complex: RNA polymerase holoenzyme with sigma factor sigB
    • Protein or peptide: DNA-directed RNA polymerase subunit alphaPolymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit alphaPolymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit betaPolymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit beta'Polymerase
    • Protein or peptide: DNA-directed RNA polymerase subunit omegaPolymerase
    • Protein or peptide: RNA polymerase sigma factor SigB

-
Supramolecule #1: RNA polymerase holoenzyme with sigma factor sigB

SupramoleculeName: RNA polymerase holoenzyme with sigma factor sigB / type: complex / ID: 1 / Chimera: Yes / Parent: 0 / Macromolecule list: all
Details: RNA polymerase holoenzyme assembled from individually expressed RNA polymerase core and sigma factor sigB
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
Molecular weightTheoretical: 800 KDa

-
Macromolecule #1: DNA-directed RNA polymerase subunit alpha

MacromoleculeName: DNA-directed RNA polymerase subunit alpha / type: protein_or_peptide / ID: 1 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
SequenceString: MLISQRPTLS EDVLTDNRSQ FVIEPLEPGF GYTLGNSLRR TLLSSIPGAA VTSIRIDGVL HEFTTVPGVK EDVTEIILNL KSLVVSSEED EPVTMYLRKQ GPGEVTAGDI VPPAGVTVHN PGMHIATLND KGKLEVELVV ERGRGYVPAV QNRASGAEIG RIPVDSIYSP ...String:
MLISQRPTLS EDVLTDNRSQ FVIEPLEPGF GYTLGNSLRR TLLSSIPGAA VTSIRIDGVL HEFTTVPGVK EDVTEIILNL KSLVVSSEED EPVTMYLRKQ GPGEVTAGDI VPPAGVTVHN PGMHIATLND KGKLEVELVV ERGRGYVPAV QNRASGAEIG RIPVDSIYSP VLKVTYKVDA TRVEQRTDFD KLILDVETKN SISPRDALAS AGKTLVELFG LARELNVEAE GIEIGPSPAE ADHIASFALP IDDLDLTVRS YNCLKREGVH TVGELVARTE SDLLDIRNFG QKSIDEVKIK LHQLGLSLKD SPPSFDPSEV AGYDVATGTW STEGAYDEQD YAETEQL

-
Macromolecule #2: DNA-directed RNA polymerase subunit alpha

MacromoleculeName: DNA-directed RNA polymerase subunit alpha / type: protein_or_peptide / ID: 2 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
SequenceString: MLISQRPTLS EDVLTDNRSQ FVIEPLEPGF GYTLGNSLRR TLLSSIPGAA VTSIRIDGVL HEFTTVPGVK EDVTEIILNL KSLVVSSEED EPVTMYLRKQ GPGEVTAGDI VPPAGVTVHN PGMHIATLND KGKLEVELVV ERGRGYVPAV QNRASGAEIG RIPVDSIYSP ...String:
MLISQRPTLS EDVLTDNRSQ FVIEPLEPGF GYTLGNSLRR TLLSSIPGAA VTSIRIDGVL HEFTTVPGVK EDVTEIILNL KSLVVSSEED EPVTMYLRKQ GPGEVTAGDI VPPAGVTVHN PGMHIATLND KGKLEVELVV ERGRGYVPAV QNRASGAEIG RIPVDSIYSP VLKVTYKVDA TRVEQRTDFD KLILDVETKN SISPRDALAS AGKTLVELFG LARELNVEAE GIEIGPSPAE ADHIASFALP IDDLDLTVRS YNCLKREGVH TVGELVARTE SDLLDIRNFG QKSIDEVKIK LHQLGLSLKD SPPSFDPSEV AGYDVATGTW STEGAYDEQD YAETEQL

-
Macromolecule #3: DNA-directed RNA polymerase subunit beta

MacromoleculeName: DNA-directed RNA polymerase subunit beta / type: protein_or_peptide / ID: 3 / Details: L2E3G4C5 -> V / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
SequenceString: MVLADSRQSK TAASPSPSRP QSSSNNSVPG APNRVSFAKL REPLEVPGLL DVQTDSFEWL IGSPRWRESA AERGDVNPVG GLEEVLYELS PIEDFSGSMS LSFSDPRFDD VKAPVDECKD KDMTYAAPLF VTAEFINNNT GEIKSQTVFM GDFPMMTEKG TFIINGTERV ...String:
MVLADSRQSK TAASPSPSRP QSSSNNSVPG APNRVSFAKL REPLEVPGLL DVQTDSFEWL IGSPRWRESA AERGDVNPVG GLEEVLYELS PIEDFSGSMS LSFSDPRFDD VKAPVDECKD KDMTYAAPLF VTAEFINNNT GEIKSQTVFM GDFPMMTEKG TFIINGTERV VVSQLVRSPG VYFDETIDKS TDKTLHSVKV IPSRGAWLEF DVDKRDTVGV RIDRKRRQPV TVLLKALGWT SEQIVERFGF SEIMRSTLEK DNTVGTDEAL LDIYRKLRPG EPPTKESAQT LLENLFFKEK RYDLARVGRY KVNKKLGLHV GEPITSSTLT EEDVVATIEY LVRLHEGQTT MTVPGGVEVP VETDDIDHFG NRRLRTVGEL IQNQIRVGMS RMERVVRERM TTQDVEAITP QTLINIRPVV AAIKEFFGTS QLSQFMDQNN PLSGLTHKRR LSALGPGGLS RERAGLEVRD VHPSHYGRMC PIETPEGPNI GLIGSLSVYA RVNPFGFIET PYRKVVDGVV SDEIVYLTAD EEDRHVVAQA NSPIDADGRF VEPRVLVRRK AGEVEYVPSS EVDYMDVSPR QMVSVATAMI PFLEHDDANR ALMGANMQRQ AVPLVRSEAP LVGTGMELRA AIDAGDVVVA EESGVIEEVS ADYITVMHDN GTRRTYRMRK FARSNHGTCA NQCPIVDAGD RVEAGQVIAD GPCTDDGEMA LGKNLLVAIM PWEGHNYEDA IILSNRLVEE DVLTSIHIEE HEIDARDTKL GAEEITRDIP NISDEVLADL DERGIVRIGA EVRDGDILVG KVTPKGETEL TPEERLLRAI FGEKAREVRD TSLKVPHGES GKVIGIRVFS REDEDELPAG VNELVRVYVA QKRKISDGDK LAGRHGNKGV IGKILPVEDM PFLADGTPVD IILNTHGVPR RMNIGQILET HLGWCAHSGW KVDAAKGVPD WAARLPDELL EAQPNAIVST PVFDGAQEAE LQGLLSCTLP NRDGDVLVDA DGKAMLFDGR SGEPFPYPVT VGYMYIMKLH HLVDDKIHAR STGPYSMITQ QPLGGKAQFG GQRFGEMECW AMQAYGAAYT LQELLTIKSD DTVGRVKVYE AIVKGENIPE PGIPESFKVL LKELQSLCLN VEVLSSDGAA IELREGEDED LERAAANLGI NLSRNESASV EDLA

-
Macromolecule #4: DNA-directed RNA polymerase subunit beta'

MacromoleculeName: DNA-directed RNA polymerase subunit beta' / type: protein_or_peptide / ID: 4 / Details: contains C-terminal 6xHis-tag / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
SequenceString: VNFFDELRIG LATAEDIRQW SYGEVKKPET INYRTLKPEK DGLFCEKIFG PTRDWECYCG KYKRVRFKGI ICERCGVEVT RAKVRRERMG HIELAAPVTH IWYFKGVPSR LGYLLDLAPK DLEKIIYFAA YVITSVDEEM RHNELSTLEA EMAVERKAVE DQRDGELEAR ...String:
VNFFDELRIG LATAEDIRQW SYGEVKKPET INYRTLKPEK DGLFCEKIFG PTRDWECYCG KYKRVRFKGI ICERCGVEVT RAKVRRERMG HIELAAPVTH IWYFKGVPSR LGYLLDLAPK DLEKIIYFAA YVITSVDEEM RHNELSTLEA EMAVERKAVE DQRDGELEAR AQKLEADLAE LEAEGAKADA RRKVRDGGER EMRQIRDRAQ RELDRLEDIW STFTKLAPKQ LIVDENLYRE LVDRYGEYFT GAMGAESIQK LIENFDIDAE AESLRDVIRN GKGQKKLRAL KRLKVVAAFQ QSGNSPMGMV LDAVPVIPPE LRPMVQLDGG RFATSDLNDL YRRVINRNNR LKRLIDLGAP EIIVNNEKRM LQESVDALFD NGRRGRPVTG PGNRPLKSLS DLLKGKQGRF RQNLLGKRVD YSGRSVIVVG PQLKLHQCGL PKLMALELFK PFVMKRLVDL NHAQNIKSAK RMVERQRPQV WDVLEEVIAE HPVLLNRAPT LHRLGIQAFE PMLVEGKAIQ LHPLVCEAFN ADFDGDQMAV HLPLSAEAQA EARILMLSSN NILSPASGRP LAMPRLDMVT GLYYLTTEVP GDTGEYQPAS GDHPETGVYS SPAEAIMAAD RGVLSVRAKI KVRLTQLRPP VEIEAELFGH SGWQPGDAWM AETTLGRVMF NELLPLGYPF VNKQMHKKVQ AAIINDLAER YPMIVVAQTV DKLKDAGFYW ATRSGVTVSM ADVLVPPRKK EILDHYEERA DKVEKQFQRG ALNHDERNEA LVEIWKEATD EVGQALREHY PDDNPIITIV DSGATGNFTQ TRTLAGMKGL VTNPKGEFIP RPVKSSFREG LTVLEYFINT HGARKGLADT ALRTADSGYL TRRLVDVSQD VIVREHDCQT ERGIVVELAE RAPDGTLIRD PYIETSAYAR TLGTDAVDEA GNVIVERGQD LGDPEIDALL AAGITQVKVR SVLTCATSTG VCATCYGRSM ATGKLVDIGE AVGIVAAQSI GEPGTQLTMR TFHQGGVGED ITGGLPRVQE LFEARVPRGK APIADVTGRV RLEDGERFYK ITIVPDDGGE EVVYDKISKR QRLRVFKHED GSERVLSDGD HVEVGQQLME GSADPHEVLR VQGPREVQIH LVREVQEVYR AQGVSIHDKH IEVIVRQMLR RVTIIDSGST EFLPGSLIDR AEFEAENRRV VAEGGEPAAG RPVLMGITKA SLATDSWLSA ASFQETTRVL TDAAINCRSD KLNGLKENVI IGKLIPAGTG INRYRNIAVQ PTEEARAAAY TIPSYEDQYY SPDFGAATGA AVPLDDYGYS DYRHHHHHH

-
Macromolecule #5: DNA-directed RNA polymerase subunit omega

MacromoleculeName: DNA-directed RNA polymerase subunit omega / type: protein_or_peptide / ID: 5 / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
SequenceString:
MSISQSDASL AAVPAVDQFD PSSGASGGYD TPLGITNPPI DELLDRVSSK YALVIYAAKR ARQINDYYNQ LGEGILEYVG PLVEPGLQEK PLSIALREIH ADLLEHTEGE

-
Macromolecule #6: RNA polymerase sigma factor SigB

MacromoleculeName: RNA polymerase sigma factor SigB / type: protein_or_peptide / ID: 6 / Details: contains N-terminal 6xHis-tag / Enantiomer: LEVO / EC number: DNA-directed RNA polymerase
Source (natural)Organism: Mycobacterium tuberculosis H37Rv (bacteria)
SequenceString: MGSSHHHHHH SSGLVPRGSH MADAPTRATT SRVDSDLDAQ SPAADLVRVY LNGIGKTALL NAAGEVELAK RIEAGLYAEH LLETRKRLGE NRKRDLAAVV RDGEAARRHL LEANLRLVVS LAKRYTGRGM PLLDLIQEGN LGLIRAMEKF DYTKGFKFST YATWWIRQAI ...String:
MGSSHHHHHH SSGLVPRGSH MADAPTRATT SRVDSDLDAQ SPAADLVRVY LNGIGKTALL NAAGEVELAK RIEAGLYAEH LLETRKRLGE NRKRDLAAVV RDGEAARRHL LEANLRLVVS LAKRYTGRGM PLLDLIQEGN LGLIRAMEKF DYTKGFKFST YATWWIRQAI TRGMADQSRT IRLPVHLVEQ VNKLARIKRE MHQHLGREAT DEELAAESGI PIDKINDLLE HSRDPVSLDM PVGSEEEAPL GDFIEDAEAM SAENAVIAEL LHTDIRSVLA TLDEREHQVI RLRFGLDDGQ PRTLDQIGKL FGLSRERVRQ IERDVMSKLR HGERADRLRS YAS

-
Experimental details

-
Structure determination

Methodcryo EM
Processingsingle particle reconstruction
Aggregation stateparticle

-
Sample preparation

Concentration1.4 mg/mL
BufferpH: 7.9
VitrificationCryogen name: ETHANE

-
Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Number real images: 3064 / Average electron dose: 49.6 e/Å2
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

-
Image processing

Particle selectionNumber selected: 254380
Startup modelType of model: NONE / Details: ab-into reconstruction
Initial angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC
Final 3D classificationSoftware - Name: cryoSPARC
Final angle assignmentType: MAXIMUM LIKELIHOOD / Software - Name: cryoSPARC
Final reconstructionNumber classes used: 1 / Applied symmetry - Point group: C1 (asymmetric) / Algorithm: FOURIER SPACE / Resolution.type: BY AUTHOR / Resolution: 4.38 Å / Resolution method: FSC 0.143 CUT-OFF / Software - Name: cryoSPARC (ver. 2.1) / Number images used: 66519
FSC plot (resolution estimation)

-
Atomic model buiding 1

Initial modelPDB ID:

7pp4

RefinementSpace: REAL / Protocol: RIGID BODY FIT

+
About Yorodumi

-
News

-
Feb 9, 2022. New format data for meta-information of EMDB entries

New format data for meta-information of EMDB entries

  • Version 3 of the EMDB header file is now the official format.
  • The previous official version 1.9 will be removed from the archive.

Related info.:EMDB header

External links:wwPDB to switch to version 3 of the EMDB data model

-
Aug 12, 2020. Covid-19 info

Covid-19 info

URL: https://pdbj.org/emnavi/covid19.php

New page: Covid-19 featured information page in EM Navigator.

Related info.:Covid-19 info / Mar 5, 2020. Novel coronavirus structure data

+
Mar 5, 2020. Novel coronavirus structure data

Novel coronavirus structure data

Related info.:Yorodumi Speices / Aug 12, 2020. Covid-19 info

External links:COVID-19 featured content - PDBj / Molecule of the Month (242)：Coronavirus Proteases

+
Jan 31, 2019. EMDB accession codes are about to change! (news from PDBe EMDB page)

EMDB accession codes are about to change! (news from PDBe EMDB page)

  • The allocation of 4 digits for EMDB accession codes will soon come to an end. Whilst these codes will remain in use, new EMDB accession codes will include an additional digit and will expand incrementally as the available range of codes is exhausted. The current 4-digit format prefixed with “EMD-” (i.e. EMD-XXXX) will advance to a 5-digit format (i.e. EMD-XXXXX), and so on. It is currently estimated that the 4-digit codes will be depleted around Spring 2019, at which point the 5-digit format will come into force.
  • The EM Navigator/Yorodumi systems omit the EMD- prefix.

Related info.:Q: What is EMD? / ID/Accession-code notation in Yorodumi/EM Navigator

External links:EMDB Accession Codes are Changing Soon! / Contact to PDBj

+
Jul 12, 2017. Major update of PDB

Major update of PDB

  • wwPDB released updated PDB data conforming to the new PDBx/mmCIF dictionary.
  • This is a major update changing the version number from 4 to 5, and with Remediation, in which all the entries are updated.
  • In this update, many items about electron microscopy experimental information are reorganized (e.g. em_software).
  • Now, EM Navigator and Yorodumi are based on the updated data.

External links:wwPDB Remediation / Enriched Model Files Conforming to OneDep Data Standards Now Available in the PDB FTP Archive

-
Yorodumi

Thousand views of thousand structures

  • Yorodumi is a browser for structure data from EMDB, PDB, SASBDB, etc.
  • This page is also the successor to EM Navigator detail page, and also detail information page/front-end page for Omokage search.
  • The word "yorodu" (or yorozu) is an old Japanese word meaning "ten thousand". "mi" (miru) is to see.

Related info.:EMDB / PDB / SASBDB / Comparison of 3 databanks / Yorodumi Search / Aug 31, 2016. New EM Navigator & Yorodumi / Yorodumi Papers / Jmol/JSmol / Function and homology information / Changes in new EM Navigator and Yorodumi

Read more