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Yorodumi- EMDB-27935: Mycobacterium tuberculosis RNAP paused elongation complex with Nu... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-27935 | |||||||||
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Title | Mycobacterium tuberculosis RNAP paused elongation complex with NusG transcription factor | |||||||||
Map data | ||||||||||
Sample |
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Keywords | RNA polymerase / transcription factor / elongation / pausing / transcription-transferase-DNA complex / TRANSCRIPTION-DNA-RNA complex | |||||||||
Function / homology | Function and homology information transcription elongation-coupled chromatin remodeling / DNA-directed RNA polymerase complex / regulation of DNA-templated transcription elongation / transcription antitermination / DNA-templated transcription termination / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / protein dimerization activity / DNA-templated transcription ...transcription elongation-coupled chromatin remodeling / DNA-directed RNA polymerase complex / regulation of DNA-templated transcription elongation / transcription antitermination / DNA-templated transcription termination / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / protein dimerization activity / DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / cytoplasm Similarity search - Function | |||||||||
Biological species | Mycobacterium tuberculosis (bacteria) / synthetic construct (others) | |||||||||
Method | single particle reconstruction / cryo EM / Resolution: 2.94 Å | |||||||||
Authors | Delbeau M / Darst SA / Campbell EA | |||||||||
Funding support | United States, 1 items
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Citation | Journal: Mol Cell / Year: 2023 Title: Structural and functional basis of the universal transcription factor NusG pro-pausing activity in Mycobacterium tuberculosis. Authors: Madeleine Delbeau / Expery O Omollo / Ruby Froom / Steven Koh / Rachel A Mooney / Mirjana Lilic / Joshua J Brewer / Jeremy Rock / Seth A Darst / Elizabeth A Campbell / Robert Landick / Abstract: Transcriptional pauses mediate regulation of RNA biogenesis. DNA-encoded pause signals trigger pausing by stabilizing RNA polymerase (RNAP) swiveling and inhibiting DNA translocation. The N-terminal ...Transcriptional pauses mediate regulation of RNA biogenesis. DNA-encoded pause signals trigger pausing by stabilizing RNA polymerase (RNAP) swiveling and inhibiting DNA translocation. The N-terminal domain (NGN) of the only universal transcription factor, NusG/Spt5, modulates pausing through contacts to RNAP and DNA. Pro-pausing NusGs enhance pauses, whereas anti-pausing NusGs suppress pauses. Little is known about pausing and NusG in the human pathogen Mycobacterium tuberculosis (Mtb). We report that MtbNusG is pro-pausing. MtbNusG captures paused, swiveled RNAP by contacts to the RNAP protrusion and nontemplate-DNA wedged between the NGN and RNAP gate loop. In contrast, anti-pausing Escherichia coli (Eco) NGN contacts the MtbRNAP gate loop, inhibiting swiveling and pausing. Using CRISPR-mediated genetics, we show that pro-pausing NGN is required for mycobacterial fitness. Our results define an essential function of mycobacterial NusG and the structural basis of pro- versus anti-pausing NusG activity, with broad implications for the function of all NusG orthologs. | |||||||||
History |
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-Structure visualization
Supplemental images |
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-Downloads & links
-EMDB archive
Map data | emd_27935.map.gz | 97.3 MB | EMDB map data format | |
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Header (meta data) | emd-27935-v30.xml emd-27935.xml | 24.4 KB 24.4 KB | Display Display | EMDB header |
Images | emd_27935.png | 41.3 KB | ||
Filedesc metadata | emd-27935.cif.gz | 8.1 KB | ||
Others | emd_27935_half_map_1.map.gz emd_27935_half_map_2.map.gz | 95.5 MB 95.5 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-27935 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-27935 | HTTPS FTP |
-Validation report
Summary document | emd_27935_validation.pdf.gz | 920.4 KB | Display | EMDB validaton report |
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Full document | emd_27935_full_validation.pdf.gz | 919.9 KB | Display | |
Data in XML | emd_27935_validation.xml.gz | 13.5 KB | Display | |
Data in CIF | emd_27935_validation.cif.gz | 15.8 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-27935 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-27935 | HTTPS FTP |
-Related structure data
Related structure data | 8e74MC 8e79C 8e82C 8e8mC 8e95C M: atomic model generated by this map C: citing same article (ref.) |
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Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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Related items in Molecule of the Month |
-Map
File | Download / File: emd_27935.map.gz / Format: CCP4 / Size: 103 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 1.0825 Å | ||||||||||||||||||||||||||||||||||||
Density |
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Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
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-Supplemental data
-Half map: #2
File | emd_27935_half_map_1.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Half map: #1
File | emd_27935_half_map_2.map | ||||||||||||
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Projections & Slices |
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Density Histograms |
-Sample components
+Entire : Mycobacterium tuberculosis RNAP paused elongation complex with Nu...
+Supramolecule #1: Mycobacterium tuberculosis RNAP paused elongation complex with Nu...
+Macromolecule #1: Transcription termination/antitermination protein NusG
+Macromolecule #2: DNA-directed RNA polymerase subunit alpha
+Macromolecule #3: DNA-directed RNA polymerase subunit beta
+Macromolecule #4: DNA-directed RNA polymerase subunit beta'
+Macromolecule #5: DNA-directed RNA polymerase subunit omega
+Macromolecule #6: DNA (54-MER)
+Macromolecule #7: DNA (54-MER)
+Macromolecule #8: RNA (42-MER)
+Macromolecule #9: ZINC ION
+Macromolecule #10: MAGNESIUM ION
-Experimental details
-Structure determination
Method | cryo EM |
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Processing | single particle reconstruction |
Aggregation state | particle |
-Sample preparation
Buffer | pH: 7.5 |
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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) / Average electron dose: 63.89 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 2.0 µm / Nominal defocus min: 1.0 µm |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
-Image processing
Startup model | Type of model: PDB ENTRY PDB model - PDB ID: |
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Final reconstruction | Resolution.type: BY AUTHOR / Resolution: 2.94 Å / Resolution method: FSC 0.143 CUT-OFF / Number images used: 264224 |
Initial angle assignment | Type: PROJECTION MATCHING |
Final angle assignment | Type: PROJECTION MATCHING |