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Open data
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Basic information
Entry | Database: EMDB / ID: EMD-21852 | |||||||||
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Title | F. tularensis RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA complex | |||||||||
![]() | F. tularensis RNAPs70-(MglA-SspA)-ppGpp-PigR-iglA DNA unsharpened map | |||||||||
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![]() | RNA polymerase complex / TRANSCRIPTION | |||||||||
Function / homology | ![]() 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 | ![]() ![]() | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 4.43 Å | |||||||||
![]() | Travis BA / Brennan RG | |||||||||
Funding support | ![]()
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![]() | ![]() 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 |
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Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
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Downloads & links
-EMDB archive
Map data | ![]() | 131 MB | ![]() | |
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Header (meta data) | ![]() ![]() | 26.8 KB 26.8 KB | Display Display | ![]() |
FSC (resolution estimation) | ![]() | 12.6 KB | Display | ![]() |
Images | ![]() | 53.5 KB | ||
Filedesc metadata | ![]() | 9 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 6wmtMC ![]() 6wegC ![]() 6wmpC ![]() 6wmrC ![]() 6wmuC M: atomic model generated by this map C: citing same article ( |
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Similar structure data |
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Links
EMDB pages | ![]() ![]() |
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Related items in Molecule of the Month |
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Map
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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
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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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![]() | single particle reconstruction |
Aggregation state | particle |
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Sample preparation
Concentration | 0.4 mg/mL |
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Buffer | pH: 8 |
Vitrification | Cryogen name: ETHANE |
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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: ![]() |
Electron optics | Illumination mode: OTHER / Imaging mode: BRIGHT FIELD / Cs: 2.7 mm |
Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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Image processing
-Atomic model buiding 1
Refinement | Space: REAL |
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Output model | ![]() PDB-6wmt: |