6fbv

Single particle cryo em structure of Mycobacterium tuberculosis RNA polymerase in complex with Fidaxomicin

Method: ELECTRON MICROSCOPY Dmax: 185.1 Å Quality: REASONABLE

1. Protein Identity and Related Structures Protein Identity & Related Structures

DNA-directed RNA polymerase subunit alpha

Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

UniProt P9WGZ1

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 6 PDB declaration: hexameric(6) Consistent with protein copy count Chain A; UniProt 1–347 Chain B; UniProt 1–347 Not recorded DNA-directed RNA polymerase subunit beta × 1 (P9WGY9) ;DNA-directed RNA polymerase subunit beta' ; × 1 (P9WGY7) DNA-directed RNA polymerase subunit omega × 1 (P9WGY5) RNA polymerase sigma factor SigA × 1 (P9WGI1) ZN ZINC ION × 2 MG MAGNESIUM ION × 1 FI8 Fidaxomicin × 1 ELECTRON MICROSCOPY cryo-EM buffer:pH 8;3.5 microliter 1 microM Mtb RNAP-Lpm and 50 microMolar Lpm in 20 mM Tris-HCl, pH 8.0, 75 mM NaCl, 5 mM MgCl2, 5 mM dithiothreitol, and 0.1% n-octyl-beta-D-glucopyranoside cryo-EM vitrification conditions:Cryogen ETHANE Resolution 3.52 Å R-free 0.331

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

50 other PDB entries and 50 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name RPOA_MYCTU
Isoform
PDB entities 1
Chains and sequence ranges Author chain A; PDBConstruct 1–347; UniProt 1–347 Author chain B; PDBConstruct 1–347; UniProt 1–347

DNA-directed RNA polymerase subunit beta

Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

UniProt P9WGY9

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 6 PDB declaration: hexameric(6) Consistent with protein copy count Chain C; UniProt 1–1178 Not recorded DNA-directed RNA polymerase subunit alpha × 2 (P9WGZ1) ;DNA-directed RNA polymerase subunit beta' ; × 1 (P9WGY7) DNA-directed RNA polymerase subunit omega × 1 (P9WGY5) RNA polymerase sigma factor SigA × 1 (P9WGI1) ZN ZINC ION × 2 MG MAGNESIUM ION × 1 FI8 Fidaxomicin × 1 ELECTRON MICROSCOPY cryo-EM buffer:pH 8;3.5 microliter 1 microM Mtb RNAP-Lpm and 50 microMolar Lpm in 20 mM Tris-HCl, pH 8.0, 75 mM NaCl, 5 mM MgCl2, 5 mM dithiothreitol, and 0.1% n-octyl-beta-D-glucopyranoside cryo-EM vitrification conditions:Cryogen ETHANE Resolution 3.52 Å R-free 0.331

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

53 other PDB entries and 54 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name RPOB_MYCTU
Isoform
PDB entities 2
Chains and sequence ranges Author chain C; PDBConstruct 1–1178; UniProt 1–1178

;DNA-directed RNA polymerase subunit beta' ;

Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

UniProt P9WGY7

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 6 PDB declaration: hexameric(6) Consistent with protein copy count Chain D; UniProt 1–1316 Not recorded DNA-directed RNA polymerase subunit alpha × 2 (P9WGZ1) DNA-directed RNA polymerase subunit beta × 1 (P9WGY9) DNA-directed RNA polymerase subunit omega × 1 (P9WGY5) RNA polymerase sigma factor SigA × 1 (P9WGI1) ZN ZINC ION × 2 MG MAGNESIUM ION × 1 FI8 Fidaxomicin × 1 ELECTRON MICROSCOPY cryo-EM buffer:pH 8;3.5 microliter 1 microM Mtb RNAP-Lpm and 50 microMolar Lpm in 20 mM Tris-HCl, pH 8.0, 75 mM NaCl, 5 mM MgCl2, 5 mM dithiothreitol, and 0.1% n-octyl-beta-D-glucopyranoside cryo-EM vitrification conditions:Cryogen ETHANE Resolution 3.52 Å R-free 0.331

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

51 other PDB entries and 51 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name RPOC_MYCTU
Isoform
PDB entities 3
Chains and sequence ranges Author chain D; PDBConstruct 1–1316; UniProt 1–1316

DNA-directed RNA polymerase subunit omega

Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

UniProt P9WGY5

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 6 PDB declaration: hexameric(6) Consistent with protein copy count Chain E; UniProt 1–110 Not recorded DNA-directed RNA polymerase subunit alpha × 2 (P9WGZ1) DNA-directed RNA polymerase subunit beta × 1 (P9WGY9) ;DNA-directed RNA polymerase subunit beta' ; × 1 (P9WGY7) RNA polymerase sigma factor SigA × 1 (P9WGI1) ZN ZINC ION × 2 MG MAGNESIUM ION × 1 FI8 Fidaxomicin × 1 ELECTRON MICROSCOPY cryo-EM buffer:pH 8;3.5 microliter 1 microM Mtb RNAP-Lpm and 50 microMolar Lpm in 20 mM Tris-HCl, pH 8.0, 75 mM NaCl, 5 mM MgCl2, 5 mM dithiothreitol, and 0.1% n-octyl-beta-D-glucopyranoside cryo-EM vitrification conditions:Cryogen ETHANE Resolution 3.52 Å R-free 0.331

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

46 other PDB entries and 46 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name RPOZ_MYCTU
Isoform
PDB entities 4
Chains and sequence ranges Author chain E; PDBConstruct 1–110; UniProt 1–110

RNA polymerase sigma factor SigA

Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)

UniProt P9WGI1

State in the Current Structure

Assembly Oligomeric State Construct Mutations and Modifications Ligands, Ions and Associated Components Method and Experimental Conditions Structure Quality
1 Protein heterocomplex Heteromer Protein × 6 PDB declaration: hexameric(6) Consistent with protein copy count Chain F; UniProt 1–528 Not recorded DNA-directed RNA polymerase subunit alpha × 2 (P9WGZ1) DNA-directed RNA polymerase subunit beta × 1 (P9WGY9) ;DNA-directed RNA polymerase subunit beta' ; × 1 (P9WGY7) DNA-directed RNA polymerase subunit omega × 1 (P9WGY5) ZN ZINC ION × 2 MG MAGNESIUM ION × 1 FI8 Fidaxomicin × 1 ELECTRON MICROSCOPY cryo-EM buffer:pH 8;3.5 microliter 1 microM Mtb RNAP-Lpm and 50 microMolar Lpm in 20 mM Tris-HCl, pH 8.0, 75 mM NaCl, 5 mM MgCl2, 5 mM dithiothreitol, and 0.1% n-octyl-beta-D-glucopyranoside cryo-EM vitrification conditions:Cryogen ETHANE Resolution 3.52 Å R-free 0.331

Other States of the Same Protein in the Database

Each row is a biological assembly of the same UniProt protein in another PDB entry. The “Difference from current entry” column identifies evidence-level differences; no tag means the currently parsed fields agree.

20 other PDB entries and 25 assemblies. Open the comparison page and filter oligomeric states

View Construct and Data Evidence
UniProt name SIGA_MYCTU
Isoform
PDB entities 5
Chains and sequence ranges Author chain F; PDBConstruct 1–528; UniProt 1–528

The page prioritizes protein identity, the current assembly, associated components, oligomeric state and cross-PDB links. Chain mapping and sequence ranges are retained as data evidence. Internal IDs, import timestamps and assembly operation expressions are maintenance fields and are not shown here.

SAXS scattering curve SAXS Profile

SAXS profile for 6fbv

P(r) Distance Distribution P(r) Distribution

P(r) distribution for 6fbv
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2. Structure Basics 2. Structure Basics

Entry ID entry_id6fbv
Deposition date deposition_date2017-12-19
Structure title titleSingle particle cryo em structure of Mycobacterium tuberculosis RNA polymerase in complex with Fidaxomicin
Keywords keywordsLipiarmycin, RNA pol, RNAP, inhibitor, drug, Clostridium difficile, ANTIBIOTIC, Tiacumicin B, CCDC 114782, transcription; TRANSCRIPTION
Experimental Method methodELECTRON MICROSCOPY

3. SAXS Parameters (CRYSOL theoretical calculation) 3. SAXS Parameters (CRYSOL)

Radius of gyration Rg (Guinier) rg_guinier50.31
Radius of gyration Rg (electron density) rg_electron49.74
Forward intensity I(0) i01884280000.00
Molecular weight molecular_weight357690.0 kDa
Excluded volume excluded_volume446730 ų
Envelope volume envelope_volume671200 ų
Hydration-shell volume shell_volume108690 ų
Envelope diameter envelope_diameter195.7
Shell Rg shell_rg56.44
Envelope Rg envelope_rg49.84
Shape Rg shape_rg49.75
Total Rg total_rg49.95
Total atoms total_atoms25143
Residues n_residues3217
Spherical-harmonic order n_harmonics20
q range q_range— – 0.5000 −1
Data points n_points101
Shell type shell_typedirectional
Solvent electron density solvent_density0.3340 e/ų
Shell contrast contrast_shell0.0300 e/ų
CRYSOL version crysol_version4.1.3

4. P(r) Distance Distribution (GNOM inversion) 4. P(r) Analysis (GNOM)

Maximum dimension Dmax dmax185.1
Rg (real space) rg_real52.85
Rg uncertainty (real space) rg_real_error0.78
I(0) (real space) i0_real1.8930e+09
I(0) uncertainty (real space) i0_real_error3.1100e+07
Rg (reciprocal space) rg_reciprocal50.35
I(0) (reciprocal space) i0_reciprocal1884000000.0000
Solution quality estimate total_estimate0.6424
Solution quality rating solution_quality REASONABLE a REASONABLE solution
P(r) peaks n_peaks1
Primary peak position r_peak_primary65.7
Skewness Skewness skewness0.656
Kurtosis Kurtosis kurtosis0.567
Angular range angular_range— – 0.1550 −1
Current regularization parameter α current_alpha0.9206
Highest regularization parameter α highest_alpha288000000.0000
Real-space data points n_real_points32
GNOM version gnom_version4.1.3
Quality Criteria quality_criteria AN1: 0.000; Oscil: 0.631; Stabil: 0.874; Sysdev: 0.000; Positv: 1.000; Valcen: 0.997; Smooth: 0.859

5. Crystallography and Experiment 5. Crystallography & Experiment

6. Entities and Polymers Entities & Polymers (9)

7. Fold Classification (SCOP + CATH) 6 domains

CATH v4.4 (6 domains)

Domain ID domain_id6fbvA01
Class class3 — Alpha Beta
Architecture architecture30 — 2-Layer Sandwich
Topology topology1360 — Gyrase A; domain 2
Homologous superfamily homologous superfamily10 — RNA polymerase, RBP11-like subunit
Domain ID domain_id6fbvA02
Class class2 — Mainly Beta
Architecture architecture170 — Beta Complex
Topology topology120 — RNA Polymerase Alpha Subunit; Chain A, domain 2
Homologous superfamily homologous superfamily12 — DNA-directed RNA polymerase, insert domain
Domain ID domain_id6fbvB01
Class class3 — Alpha Beta
Architecture architecture30 — 2-Layer Sandwich
Topology topology1360 — Gyrase A; domain 2
Homologous superfamily homologous superfamily10 — RNA polymerase, RBP11-like subunit
Domain ID domain_id6fbvB02
Class class2 — Mainly Beta
Architecture architecture170 — Beta Complex
Topology topology120 — RNA Polymerase Alpha Subunit; Chain A, domain 2
Homologous superfamily homologous superfamily12 — DNA-directed RNA polymerase, insert domain
Domain ID domain_id6fbvD01
Class class1 — Mainly Alpha
Architecture architecture10 — Orthogonal Bundle
Topology topology274 — Enzyme I; Chain A, domain 2
Homologous superfamily homologous superfamily100 — RNA polymerase Rpb1, domain 3
Domain ID domain_id6fbvD02
Class class1 — Mainly Alpha
Architecture architecture10 — Orthogonal Bundle
Topology topology132 — Topoisomerase I; Chain A, domain 4
Homologous superfamily homologous superfamily30 — RNA polymerase Rpb1 funnel domain

8. Citations (1)

9. Files and Curves (10)