+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-21852 | |||||||||
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Title | F. tularensis RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA complex | |||||||||
Map data | F. tularensis RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA unsharpened map | |||||||||
Sample |
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Keywords | RNA polymerase complex / TRANSCRIPTION | |||||||||
Function / homology | Function and homology information sigma factor activity / DNA-directed RNA polymerase complex / glutathione metabolic process / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / transferase activity / protein dimerization activity / DNA-templated transcription ...sigma factor activity / DNA-directed RNA polymerase complex / glutathione metabolic process / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / transferase activity / protein dimerization activity / DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / cytoplasm Similarity search - Function | |||||||||
Biological species | Francisella tularensis subsp. holarctica LVS (bacteria) / Francisella tularensis subsp. holarctica (strain LVS) (bacteria) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.43 Å | |||||||||
Authors | Travis BA / Brennan RG | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Mol Cell / Year: 2021 Title: Structural Basis for Virulence Activation of Francisella tularensis. Authors: Brady A Travis / Kathryn M Ramsey / Samantha M Prezioso / Thomas Tallo / Jamie M Wandzilak / Allen Hsu / Mario Borgnia / Alberto Bartesaghi / Simon L Dove / Richard G Brennan / Maria A Schumacher / Abstract: The bacterium Francisella tularensis (Ft) is one of the most infectious agents known. Ft virulence is controlled by a unique combination of transcription regulators: the MglA-SspA heterodimer, PigR, ...The bacterium Francisella tularensis (Ft) is one of the most infectious agents known. Ft virulence is controlled by a unique combination of transcription regulators: the MglA-SspA heterodimer, PigR, and the stress signal, ppGpp. MglA-SspA assembles with the σ-associated RNAP holoenzyme (RNAPσ), forming a virulence-specialized polymerase. These factors activate Francisella pathogenicity island (FPI) gene expression, which is required for virulence, but the mechanism is unknown. Here we report FtRNAPσ-promoter-DNA, FtRNAPσ-(MglA-SspA)-promoter DNA, and FtRNAPσ-(MglA-SspA)-ppGpp-PigR-promoter DNA cryo-EM structures. Structural and genetic analyses show MglA-SspA facilitates σ binding to DNA to regulate virulence and virulence-enhancing genes. Our Escherichia coli RNAPσhomodimeric EcSspA structure suggests this is a general SspA-transcription regulation mechanism. Strikingly, our FtRNAPσ-(MglA-SspA)-ppGpp-PigR-DNA structure reveals ppGpp binding to MglA-SspA tethers PigR to promoters. PigR in turn recruits FtRNAP αCTDs to DNA UP elements. Thus, these studies unveil a unique mechanism for Ft pathogenesis involving a virulence-specialized RNAP that employs two (MglA-SspA)-based strategies to activate virulence genes. | |||||||||
History |
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-Structure visualization
Movie |
Movie viewer |
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Structure viewer | EM map: SurfViewMolmilJmol/JSmol |
Supplemental images |
-Downloads & links
-EMDB archive
Map data | emd_21852.map.gz | 131 MB | EMDB map data format | |
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Header (meta data) | emd-21852-v30.xml emd-21852.xml | 26.8 KB 26.8 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_21852_fsc.xml | 12.6 KB | Display | FSC data file |
Images | emd_21852.png | 53.5 KB | ||
Filedesc metadata | emd-21852.cif.gz | 9 KB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-21852 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-21852 | HTTPS FTP |
-Validation report
Summary document | emd_21852_validation.pdf.gz | 535.6 KB | Display | EMDB validaton report |
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Full document | emd_21852_full_validation.pdf.gz | 535.1 KB | Display | |
Data in XML | emd_21852_validation.xml.gz | 12.9 KB | Display | |
Data in CIF | emd_21852_validation.cif.gz | 17.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21852 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-21852 | HTTPS FTP |
-Related structure data
Related structure data | 6wmtMC 6wegC 6wmpC 6wmrC 6wmuC M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_21852.map.gz / Format: CCP4 / Size: 166.4 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Annotation | F. tularensis RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA unsharpened map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.07 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
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-Supplemental data
-Sample components
+Entire : Francisella RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA complex
+Supramolecule #1: Francisella RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA complex
+Macromolecule #1: DNA-directed RNA polymerase subunit omega
+Macromolecule #2: DNA-directed RNA polymerase subunit alpha 1
+Macromolecule #3: DNA-directed RNA polymerase subunit alpha 2
+Macromolecule #4: DNA-directed RNA polymerase subunit beta
+Macromolecule #5: DNA-directed RNA polymerase subunit beta'
+Macromolecule #9: aCTDs
+Macromolecule #10: Macrophage growth locus, subunit A
+Macromolecule #11: Glutathione S-transferase
+Macromolecule #13: RNA polymerase sigma factor RpoD
+Macromolecule #6: DNA NT-strand
+Macromolecule #7: DNA T-strand
+Macromolecule #8: DNA T-strand downstream
+Macromolecule #12: DNA NT-strand downstream
+Macromolecule #14: MAGNESIUM ION
+Macromolecule #15: ZINC ION
+Macromolecule #16: GUANOSINE-5',3'-TETRAPHOSPHATE
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Concentration | 0.4 mg/mL |
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Buffer | pH: 8 |
Vitrification | Cryogen name: ETHANE |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
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Image recording | Film or detector model: GATAN K3 (6k x 4k) / Detector mode: COUNTING / Average electron dose: 60.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Refinement | Space: REAL |
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Output model | PDB-6wmt: |