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Open data
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Basic information
| Entry | Database: EMDB / ID: EMD-7323 | |||||||||
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| Title | Mycobacterium tuberculosis RNAP Holo/RbpA/Fidaxomicin | |||||||||
Map data | primary map | |||||||||
Sample |
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Keywords | initiation / antibiotic / switch region inhibitor / TRANSCRIPTION / transcription-antibiotic complex | |||||||||
| Function / homology | Function and homology informationresponse to water / bacterial-type RNA polymerase holo enzyme binding / Antimicrobial action and antimicrobial resistance in Mtb / sigma factor activity / bacterial-type RNA polymerase core enzyme binding / cytosolic DNA-directed RNA polymerase complex / DNA-directed RNA polymerase complex / peptidoglycan-based cell wall / DNA-templated transcription initiation / ribonucleoside binding ...response to water / bacterial-type RNA polymerase holo enzyme binding / Antimicrobial action and antimicrobial resistance in Mtb / sigma factor activity / bacterial-type RNA polymerase core enzyme binding / cytosolic DNA-directed RNA polymerase complex / DNA-directed RNA polymerase complex / peptidoglycan-based cell wall / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed RNA polymerase / DNA-directed RNA polymerase activity / nucleic acid binding / protein dimerization activity / response to antibiotic / DNA-templated transcription / positive regulation of DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / plasma membrane / cytoplasm / cytosol Similarity search - Function | |||||||||
| Biological species | ![]() | |||||||||
| Method | single particle reconstruction / cryo EM / Resolution: 5.15 Å | |||||||||
Authors | Darst SA / Campbell EA | |||||||||
Citation | Journal: Elife / Year: 2018Title: Fidaxomicin jams RNA polymerase motions needed for initiation via RbpA contacts. Authors: Hande Boyaci / James Chen / Mirjana Lilic / Margaret Palka / Rachel Anne Mooney / Robert Landick / Seth A Darst / Elizabeth A Campbell / ![]() Abstract: Fidaxomicin (Fdx) is an antimicrobial RNA polymerase (RNAP) inhibitor highly effective against RNAP in vitro, but clinical use of Fdx is limited to treating intestinal infections due to poor ...Fidaxomicin (Fdx) is an antimicrobial RNA polymerase (RNAP) inhibitor highly effective against RNAP in vitro, but clinical use of Fdx is limited to treating intestinal infections due to poor absorption. To identify the structural determinants of Fdx binding to RNAP, we determined the 3.4 Å cryo-electron microscopy structure of a complete RNAP holoenzyme in complex with Fdx. We find that the actinobacteria general transcription factor RbpA contacts fidaxomycin, explaining its strong effect on . Additional structures define conformational states of RNAP between the free apo-holoenzyme and the promoter-engaged open complex ready for transcription. The results establish that Fdx acts like a doorstop to jam the enzyme in an open state, preventing the motions necessary to secure promoter DNA in the active site. Our results provide a structural platform to guide development of anti-tuberculosis antimicrobials based on the Fdx binding pocket. #1: Journal: To Be PublishedTitle: Structure of Mycobacterium Tuberculosis RNAP Holo Enzyme/RbpA in closed clamp conformation Authors: Darst SA / Campbell EA / Boyaci Selcuk H / Chen J / Lilic M | |||||||||
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Structure visualization
| Movie |
Movie viewer |
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| Structure viewer | EM map: SurfView Molmil Jmol/JSmol |
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Downloads & links
-EMDB archive
| Map data | emd_7323.map.gz | 38 MB | EMDB map data format | |
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| Header (meta data) | emd-7323-v30.xml emd-7323.xml | 21.4 KB 21.4 KB | Display Display | EMDB header |
| Images | emd_7323.png | 121.3 KB | ||
| Filedesc metadata | emd-7323.cif.gz | 8.7 KB | ||
| Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-7323 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-7323 | HTTPS FTP |
-Validation report
| Summary document | emd_7323_validation.pdf.gz | 523.7 KB | Display | EMDB validaton report |
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| Full document | emd_7323_full_validation.pdf.gz | 523.3 KB | Display | |
| Data in XML | emd_7323_validation.xml.gz | 5.1 KB | Display | |
| Data in CIF | emd_7323_validation.cif.gz | 5.9 KB | Display | |
| Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-7323 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-7323 | HTTPS FTP |
-Related structure data
| Related structure data | ![]() 6c06MC ![]() 7319C ![]() 7320C ![]() 7322C ![]() 6bzoC ![]() 6c04C ![]() 6c05C 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 | EMDB (EBI/PDBe) / EMDataResource |
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| Related items in Molecule of the Month |
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Map
| File | Download / File: emd_7323.map.gz / Format: CCP4 / Size: 40.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Annotation | primary map | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Projections & slices | Image control
Images are generated by Spider. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Voxel size | X=Y=Z: 1.5 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 : Mycobacterium tuberculosis RNAP Holo with RbpA and Fidaxomicin
+Supramolecule #1: Mycobacterium tuberculosis RNAP Holo with RbpA and Fidaxomicin
+Macromolecule #1: DNA-directed RNA polymerase subunit alpha
+Macromolecule #2: DNA-directed RNA polymerase subunit beta
+Macromolecule #3: DNA-directed RNA polymerase subunit beta'
+Macromolecule #4: DNA-directed RNA polymerase subunit omega
+Macromolecule #5: RNA polymerase sigma factor SigA
+Macromolecule #6: RNA polymerase-binding protein RbpA
+Macromolecule #7: ZINC ION
+Macromolecule #8: MAGNESIUM ION
+Macromolecule #9: Fidaxomicin
-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: 8 |
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| Vitrification | Cryogen name: ETHANE |
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Electron microscopy
| Microscope | FEI TALOS ARCTICA |
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| Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Average electron dose: 6.7 e/Å2 |
| Electron beam | Acceleration voltage: 200 kV / Electron source: FIELD EMISSION GUN |
| Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD |
| Experimental equipment | ![]() Model: Talos Arctica / Image courtesy: FEI Company |
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