[English] 日本語
Yorodumi
- PDB-6c06: Mycobacterium tuberculosis RNAP Holo/RbpA/Fidaxomicin -

+
Open data


ID or keywords:

Loading...

-
Basic information

Entry
Database: PDB / ID: 6c06
TitleMycobacterium tuberculosis RNAP Holo/RbpA/Fidaxomicin
Components
  • (DNA-directed RNA polymerase subunit ...Polymerase) x 4
  • RNA polymerase sigma factor SigA
  • RNA polymerase-binding protein RbpA
Keywordstranscription/antibiotic / initiation / antibiotic / switch region inhibitor / TRANSCRIPTION / transcription-antibiotic complex
Function / homology
Function and homology information


bacterial-type RNA polymerase holo enzyme binding / response to water / bacterial-type RNA polymerase core enzyme binding / transcription initiation from bacterial-type RNA polymerase promoter / sigma factor activity / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / cell wall / transcription, DNA-templated ...bacterial-type RNA polymerase holo enzyme binding / response to water / bacterial-type RNA polymerase core enzyme binding / transcription initiation from bacterial-type RNA polymerase promoter / sigma factor activity / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / cell wall / transcription, DNA-templated / protein dimerization activity / pathogenesis / DNA-binding transcription factor activity / response to antibiotic / positive regulation of transcription, DNA-templated / magnesium ion binding / DNA binding / zinc ion binding / plasma membrane / cytosol / cytoplasm
RNA polymerase-binding protein / RNA polymerase sigma factor RpoD / RNA polymerase Rpb1, funnel domain superfamily / RNA polymerase Rpb2, domain 2 superfamily / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain superfamily / DNA-directed RNA polymerase, insert domain superfamily / RNA polymerase, RBP11-like subunit / Winged helix-like DNA-binding domain superfamily / RPB6/omega subunit-like superfamily / RNA polymerase-binding protein RbpA ...RNA polymerase-binding protein / RNA polymerase sigma factor RpoD / RNA polymerase Rpb1, funnel domain superfamily / RNA polymerase Rpb2, domain 2 superfamily / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain superfamily / DNA-directed RNA polymerase, insert domain superfamily / RNA polymerase, RBP11-like subunit / Winged helix-like DNA-binding domain superfamily / RPB6/omega subunit-like superfamily / RNA polymerase-binding protein RbpA / RNA polymerase Rpb1, domain 3 superfamily / DNA-directed RNA polymerase, beta subunit, external 1 domain / DNA-directed RNA polymerase, subunit 2 / RNA polymerase Rpb2, OB-fold / RNA polymerase sigma-70 like domain / RNA polymerase sigma factor, region 3/4-like / RNA polymerase sigma factor RpoD, C-terminal / DNA-directed RNA polymerase, subunit beta-prime / RNA polymerase subunit, RPB6/omega / RbpA superfamily / DNA-directed RNA polymerase, beta subunit, external 1 domain superfamily / DNA-directed RNA polymerase, RpoA/D/Rpb3-type / RNA polymerase Rpb1, domain 3 / Sigma-70, region 4 / Sigma-70 region 2 / Sigma-70 region 3 / Sigma-70 factor, region 1.2 / RNA polymerase Rpb6 / RNA polymerase Rpb1, domain 4 / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 1 / RNA polymerase Rpb1, domain 2 / RNA polymerase Rpb3/RpoA insert domain / RNA polymerase beta subunit external 1 domain / RNA polymerase Rpb2, domain 3 / RNA polymerase beta subunit / RNA polymerase Rpb2, domain 2 / RNA polymerase Rpb2, domain 7 / RNA polymerase Rpb2, domain 6 / Bacterial RNA polymerase, alpha chain C terminal domain / RNA polymerase Rpb3/Rpb11 dimerisation domain / DNA-directed RNA polymerase, alpha subunit / RNA polymerase sigma factor, region 2 / DNA-directed RNA polymerase, insert domain / DNA-directed RNA polymerase, subunit 2, hybrid-binding domain / RNA polymerase, alpha subunit, C-terminal / RNA polymerase, alpha subunit / RNA polymerase sigma-70 / DNA-directed RNA polymerase, omega subunit / RNA polymerase, subunit omega/K/RPB6 / RNA polymerase, N-terminal / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, domain 1 / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 4 / RNA polymerase, beta subunit, conserved site / RNA polymerase sigma-70 region 3 / DNA-directed RNA polymerase beta subunit, bacterial-type / RNA polymerase sigma-70 region 1.2 / RNA polymerase Rpb2, domain 3 / RNA polymerase, beta subunit, protrusion / RNA polymerase Rpb2, domain 2 / RNA polymerase Rpb2, domain 7 / RNA polymerase sigma-70 region 4 / RNA polymerase sigma-70 region 2
RNA polymerase sigma factor SigA / RNA polymerase-binding protein RbpA / DNA-directed RNA polymerase subunit alpha / DNA-directed RNA polymerase subunit beta' / DNA-directed RNA polymerase subunit beta / DNA-directed RNA polymerase subunit beta' / RNA polymerase sigma factor SigA / DNA-directed RNA polymerase subunit omega / DNA-directed RNA polymerase subunit beta / RNA polymerase-binding protein RbpA ...RNA polymerase sigma factor SigA / RNA polymerase-binding protein RbpA / DNA-directed RNA polymerase subunit alpha / DNA-directed RNA polymerase subunit beta' / DNA-directed RNA polymerase subunit beta / DNA-directed RNA polymerase subunit beta' / RNA polymerase sigma factor SigA / DNA-directed RNA polymerase subunit omega / DNA-directed RNA polymerase subunit beta / RNA polymerase-binding protein RbpA / DNA-directed RNA polymerase subunit alpha / DNA-directed RNA polymerase subunit omega
Biological speciesMycobacterium tuberculosis (bacteria)
MethodELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 5.15 Å
AuthorsDarst, S.A. / Campbell, E.A. / Boyaci Selcuk, H. / Chen, J. / Lilic, M.
Citation
Journal: Elife / Year: 2018
Title: Fidaxomicin jams RNA polymerase motions needed for initiation via RbpA contacts.
Authors: Hande Boyaci / James Chen / Mirjana Lilic / Margaret Palka / Rachel Anne Mooney / Robert Landick / Seth A Darst / Elizabeth A Campbell /
Abstract: Fidaxomicin (Fdx) is an antimicrobial RNA polymerase (RNAP) inhibitor highly effective against RNAP in vitro, but clinical use of Fdx is limited to treating intestinal infections due to poor ...Fidaxomicin (Fdx) is an antimicrobial RNA polymerase (RNAP) inhibitor highly effective against RNAP in vitro, but clinical use of Fdx is limited to treating intestinal infections due to poor absorption. To identify the structural determinants of Fdx binding to RNAP, we determined the 3.4 Å cryo-electron microscopy structure of a complete RNAP holoenzyme in complex with Fdx. We find that the actinobacteria general transcription factor RbpA contacts fidaxomycin, explaining its strong effect on . Additional structures define conformational states of RNAP between the free apo-holoenzyme and the promoter-engaged open complex ready for transcription. The results establish that Fdx acts like a doorstop to jam the enzyme in an open state, preventing the motions necessary to secure promoter DNA in the active site. Our results provide a structural platform to guide development of anti-tuberculosis antimicrobials based on the Fdx binding pocket.
#1: Journal: To Be Published
Title: Structure of Mycobacterium Tuberculosis RNAP Holo Enzyme/RbpA in closed clamp conformation
Authors: Darst, S.A. / Campbell, E.A. / Boyaci Selcuk, H. / Chen, J. / Lilic, M.
Validation Report
SummaryFull reportAbout validation report
History
DepositionDec 27, 2017Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 28, 2018Provider: repository / Type: Initial release

-
Structure visualization

Movie
  • Deposited structure unit
  • Imaged by Jmol
  • Download
  • Superimposition on EM map
  • EMDB-7323
  • Imaged by UCSF Chimera
  • Download
Movie viewer
Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

-
Assembly

Deposited unit
A: DNA-directed RNA polymerase subunit alpha
B: DNA-directed RNA polymerase subunit alpha
C: DNA-directed RNA polymerase subunit beta
D: DNA-directed RNA polymerase subunit beta'
E: DNA-directed RNA polymerase subunit omega
F: RNA polymerase sigma factor SigA
J: RNA polymerase-binding protein RbpA
hetero molecules


Theoretical massNumber of molelcules
Total (without water)437,91711
Polymers436,7047
Non-polymers1,2134
Water0
1


TypeNameSymmetry operationNumber
identity operation1_5551

-
Components

-
DNA-directed RNA polymerase subunit ... , 4 types, 5 molecules ABCDE

#1: Protein DNA-directed RNA polymerase subunit alpha / Polymerase / RNAP subunit alpha / RNA polymerase subunit alpha / Transcriptase subunit alpha


Mass: 37745.328 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria)
Gene: rpoA, rpoA_1, CLD25_18970, ERS007661_01801, ERS007663_01917, ERS007665_01054, ERS007670_00938, ERS007672_03180, ERS007679_01858, ERS007688_02121, ERS007703_04392, ERS007720_00529, ERS007722_ ...Gene: rpoA, rpoA_1, CLD25_18970, ERS007661_01801, ERS007663_01917, ERS007665_01054, ERS007670_00938, ERS007672_03180, ERS007679_01858, ERS007688_02121, ERS007703_04392, ERS007720_00529, ERS007722_04215, ERS023446_02770, ERS024213_02470, ERS027644_00781, ERS027646_02177, ERS027652_00409, ERS027653_00362, ERS027654_02191, ERS027656_00886, ERS027659_01235, ERS027661_02549, ERS027666_01143, ERS124361_03550, SAMEA2682864_03600, SAMEA2683035_02863
Production host: Escherichia coli (E. coli)
References: UniProt: A0A045J8T1, UniProt: P9WGZ1*PLUS, DNA-directed RNA polymerase
#2: Protein DNA-directed RNA polymerase subunit beta / Polymerase / RNAP subunit beta / RNA polymerase subunit beta / Transcriptase subunit beta


Mass: 130198.922 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria) / Gene: rpoB, SAMEA2682864_01701 / Production host: Escherichia coli (E. coli)
References: UniProt: V9Z879, UniProt: P9WGY9*PLUS, DNA-directed RNA polymerase
#3: Protein DNA-directed RNA polymerase subunit beta' / Polymerase / RNAP subunit beta' / RNA polymerase subunit beta' / Transcriptase subunit beta'


Mass: 148074.094 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria)
Gene: rpoC, rpoC_1, CLD25_03785, ERS007665_00591, ERS023446_00410, ERS031537_00289, ERS124361_01694, SAMEA2682864_01702
Production host: Escherichia coli (E. coli)
References: UniProt: A0A045J9E2, UniProt: P9WGY7*PLUS, DNA-directed RNA polymerase
#4: Protein DNA-directed RNA polymerase subunit omega / Polymerase / RNAP omega subunit / RNA polymerase omega subunit / Transcriptase subunit omega


Mass: 11776.996 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria)
Gene: rpoZ, ERS007672_03979, ERS007703_04032, ERS007720_04749, ERS027652_00548, ERS027654_02543, ERS027656_03959, ERS124361_02246
Production host: Escherichia coli (E. coli)
References: UniProt: A0A0T9N9K3, UniProt: P9WGY5*PLUS, DNA-directed RNA polymerase

-
Protein , 2 types, 2 molecules FJ

#5: Protein RNA polymerase sigma factor SigA


Mass: 58169.477 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria)
Gene: sigA, hrdB_2, CLD25_14885, ERS007663_03246, ERS007679_00877, ERS007720_03380, ERS007722_02638, ERS023446_01203, ERS024213_00371, ERS024276_00878, ERS027644_01437, ERS027646_01351, ERS027653_ ...Gene: sigA, hrdB_2, CLD25_14885, ERS007663_03246, ERS007679_00877, ERS007720_03380, ERS007722_02638, ERS023446_01203, ERS024213_00371, ERS024276_00878, ERS027644_01437, ERS027646_01351, ERS027653_01295, ERS027661_00252, ERS124361_02215, SAMEA2682864_02073, SAMEA2683035_01696
Production host: Escherichia coli (E. coli) / References: UniProt: A0A045HD00, UniProt: P9WGI1*PLUS
#6: Protein RNA polymerase-binding protein RbpA


Mass: 12993.695 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Mycobacterium tuberculosis (bacteria)
Gene: rbpA, CLD25_11245, ERS007670_01223, ERS007672_02518, ERS007679_02287, ERS007681_03389, ERS007688_02707, ERS007703_03544, ERS007720_03893, ERS007722_02881, ERS007731_02971, ERS007739_00749, ...Gene: rbpA, CLD25_11245, ERS007670_01223, ERS007672_02518, ERS007679_02287, ERS007681_03389, ERS007688_02707, ERS007703_03544, ERS007720_03893, ERS007722_02881, ERS007731_02971, ERS007739_00749, ERS007741_03837, ERS023446_04025, ERS024213_00234, ERS024276_03708, ERS027644_04191, ERS027646_02841, ERS027652_02278, ERS027653_01824, ERS027654_04301, ERS027656_03007, ERS027659_00863, ERS027661_00907, ERS027666_04473, ERS031537_02569, ERS124361_01290, SAMEA2682864_00634, SAMEA2683035_00392
Production host: Escherichia coli (E. coli) / References: UniProt: A0A045IP01, UniProt: P9WHJ5*PLUS

-
Non-polymers , 3 types, 4 molecules

#7: Chemical ChemComp-ZN / ZINC ION / Zinc


Mass: 65.409 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Zn
#8: Chemical ChemComp-MG / MAGNESIUM ION / Magnesium


Mass: 24.305 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: Mg
#9: Chemical ChemComp-FI8 / Fidaxomicin / Fidaxomicin


Mass: 1058.039 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C52H74Cl2O18 / Comment: antibiotic*YM

-
Experimental details

-
Experiment

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

-
Sample preparation

ComponentName: Mycobacterium tuberculosis RNAP Holo with RbpA and Fidaxomicin
Type: COMPLEX / Entity ID: #1-#6 / Source: RECOMBINANT
Source (natural)Organism: Mycobacterium tuberculosis (bacteria)
Source (recombinant)Organism: Escherichia coli (E. coli)
Buffer solutionpH: 8
SpecimenEmbedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES
VitrificationCryogen name: ETHANE

-
Electron microscopy imaging

Experimental equipment
Model: Talos Arctica / Image courtesy: FEI Company
MicroscopyModel: FEI TALOS ARCTICA
Electron gunElectron source: FIELD EMISSION GUN / Accelerating voltage: 200 kV / Illumination mode: FLOOD BEAM
Electron lensMode: BRIGHT FIELDBright-field microscopy
Image recordingElectron dose: 6.7 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k)

-
Processing

SoftwareName: PHENIX / Version: 1.11.1_2575: / Classification: refinement
CTF correctionType: PHASE FLIPPING AND AMPLITUDE CORRECTION
3D reconstructionResolution: 5.15 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 171547 / Symmetry type: POINT
Refine LS restraints
Refinement-IDTypeDev idealNumber
ELECTRON MICROSCOPYf_bond_d0.01126608
ELECTRON MICROSCOPYf_angle_d1.40936115
ELECTRON MICROSCOPYf_dihedral_angle_d10.85816258
ELECTRON MICROSCOPYf_chiral_restr0.0714099
ELECTRON MICROSCOPYf_plane_restr0.0094740

+
About Yorodumi

-
News

-
Aug 12, 2020. New: Covid-19 info

New: Covid-19 info

  • 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. New: 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. (see PDBe EMDB page)
  • 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 at PDBe / 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. See below links for details.
  • 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

+
Jun 16, 2017. Omokage search with filter

Omokage search with filter

  • Result of Omokage search can be filtered by keywords and the database types

Related info.:Omokage search

Read more

-
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.

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