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Yorodumi- EMDB-18650: Mycobacterium smegnatis RNAP open promoter complex with SigmaA an... -
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Open data
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Basic information
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| Title | Mycobacterium smegnatis RNAP open promoter complex with SigmaA and RbpA | ||||||||||||||||||
Map data | Loc-Scale filtered | ||||||||||||||||||
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Keywords | HelD / RNA polymerase / transcription initiation / Mycobacteria / TRANSCRIPTION / rifampicin resistance | ||||||||||||||||||
| Function / homology | Function and homology informationsigma factor activity / bacterial-type RNA polymerase core enzyme binding / DNA-directed RNA polymerase complex / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / protein dimerization activity / response to antibiotic / DNA-templated transcription ...sigma factor activity / bacterial-type RNA polymerase core enzyme binding / DNA-directed RNA polymerase complex / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / protein dimerization activity / response to antibiotic / DNA-templated transcription / positive regulation of DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / cytoplasm Similarity search - Function | ||||||||||||||||||
| Biological species | Mycolicibacterium smegmatis MC2 155 (bacteria) | ||||||||||||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 3.09 Å | ||||||||||||||||||
Authors | Koval T / Krasny L / Dohnalek J / Kouba T | ||||||||||||||||||
| Funding support | Czech Republic, European Union, 5 items
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Citation | Journal: Nat Commun / Year: 2024Title: Mycobacterial HelD connects RNA polymerase recycling with transcription initiation. Authors: Tomáš Kovaľ / Nabajyoti Borah / Petra Sudzinová / Barbora Brezovská / Hana Šanderová / Viola Vaňková Hausnerová / Alena Křenková / Martin Hubálek / Mária Trundová / Kristýna ...Authors: Tomáš Kovaľ / Nabajyoti Borah / Petra Sudzinová / Barbora Brezovská / Hana Šanderová / Viola Vaňková Hausnerová / Alena Křenková / Martin Hubálek / Mária Trundová / Kristýna Adámková / Jarmila Dušková / Marek Schwarz / Jana Wiedermannová / Jan Dohnálek / Libor Krásný / Tomáš Kouba / ![]() Abstract: Mycobacterial HelD is a transcription factor that recycles stalled RNAP by dissociating it from nucleic acids and, if present, from the antibiotic rifampicin. The rescued RNAP, however, must ...Mycobacterial HelD is a transcription factor that recycles stalled RNAP by dissociating it from nucleic acids and, if present, from the antibiotic rifampicin. The rescued RNAP, however, must disengage from HelD to participate in subsequent rounds of transcription. The mechanism of release is unknown. We show that HelD from Mycobacterium smegmatis forms a complex with RNAP associated with the primary sigma factor σ and transcription factor RbpA but not CarD. We solve several structures of RNAP-σ-RbpA-HelD without and with promoter DNA. These snapshots capture HelD during transcription initiation, describing mechanistic aspects of HelD release from RNAP and its protective effect against rifampicin. Biochemical evidence supports these findings, defines the role of ATP binding and hydrolysis by HelD in the process, and confirms the rifampicin-protective effect of HelD. Collectively, these results show that when HelD is present during transcription initiation, the process is protected from rifampicin until the last possible moment. | ||||||||||||||||||
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Structure visualization
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Downloads & links
-EMDB archive
| Map data | emd_18650.map.gz | 88.1 MB | EMDB map data format | |
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| Header (meta data) | emd-18650-v30.xml emd-18650.xml | 32.1 KB 32.1 KB | Display Display | EMDB header |
| FSC (resolution estimation) | emd_18650_fsc.xml | 12.8 KB | Display | FSC data file |
| Images | emd_18650.png | 101.9 KB | ||
| Masks | emd_18650_msk_1.map | 178 MB | Mask map | |
| Filedesc metadata | emd-18650.cif.gz | 9.6 KB | ||
| Others | emd_18650_additional_1.map.gz emd_18650_half_map_1.map.gz emd_18650_half_map_2.map.gz | 165.7 MB 141.4 MB 140.8 MB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-18650 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-18650 | HTTPS FTP |
-Validation report
| Summary document | emd_18650_validation.pdf.gz | 844.1 KB | Display | EMDB validaton report |
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| Full document | emd_18650_full_validation.pdf.gz | 843.7 KB | Display | |
| Data in XML | emd_18650_validation.xml.gz | 19.8 KB | Display | |
| Data in CIF | emd_18650_validation.cif.gz | 25.6 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18650 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-18650 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 8qtiMC ![]() 8q3iC ![]() 8qn8C ![]() 8qu6C ![]() 8r2mC ![]() 8r3mC ![]() 8r6pC ![]() 8r6rC M: atomic model generated by this map C: citing same article ( |
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| Similar structure data | Similarity search - Function & homology F&H Search |
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Links
| EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_18650.map.gz / Format: CCP4 / Size: 178 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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| Annotation | Loc-Scale filtered | ||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 0.8336 Å | ||||||||||||||||||||||||||||||||||||
| Density |
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| Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||
| Details | EMDB XML:
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-Supplemental data
-Mask #1
| File | emd_18650_msk_1.map | ||||||||||||
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-Additional map: RELION post-processed
| File | emd_18650_additional_1.map | ||||||||||||
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| Annotation | RELION post-processed | ||||||||||||
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-Half map: #2
| File | emd_18650_half_map_1.map | ||||||||||||
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-Half map: #1
| File | emd_18650_half_map_2.map | ||||||||||||
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Sample components
+Entire : Mycobacterium smegnatis RNAP open promoter complex with SigmaA an...
+Supramolecule #1: Mycobacterium smegnatis RNAP open promoter complex with SigmaA an...
+Macromolecule #1: DNA-directed RNA polymerase subunit alpha
+Macromolecule #2: DNA-directed RNA polymerase subunit beta
+Macromolecule #3: RNA polymerase sigma factor SigA
+Macromolecule #5: DNA-directed RNA polymerase subunit beta'
+Macromolecule #6: DNA-directed RNA polymerase subunit omega
+Macromolecule #7: RNA polymerase-binding protein RbpA
+Macromolecule #4: DNA 50-mer non-template strand
+Macromolecule #8: DNA 50-mer template strand
+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 |
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Sample preparation
| Buffer | pH: 7.4 |
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| 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 BIOQUANTUM (6k x 4k) / Average exposure time: 2.0 sec. / Average electron dose: 40.0 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: 2.9 µm / Nominal defocus min: 0.5 µm |
| Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
| Experimental equipment | ![]() Model: Titan Krios / Image courtesy: FEI Company |
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About Yorodumi



Keywords
Mycolicibacterium smegmatis MC2 155 (bacteria)
Authors
Czech Republic, European Union, 5 items
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Processing
FIELD EMISSION GUN


