|Entry||Database: EMDB / ID: EMD-4510|
|Title||Cryo em structure of the Listeria stressosome|
|Biological species||Listeria monocytogenes (bacteria) / Listeria monocytogenes EGD-E (bacteria)|
|Method||single particle reconstruction / cryo EM / Resolution: 3.37 Å|
|Authors||Williams AH / Redzej A|
|Citation||Journal: Nat Commun / Year: 2019|
Title: The cryo-electron microscopy supramolecular structure of the bacterial stressosome unveils its mechanism of activation.
Authors: Allison H Williams / Adam Redzej / Nathalie Rolhion / Tiago R D Costa / Aline Rifflet / Gabriel Waksman / Pascale Cossart /
Abstract: How the stressosome, the epicenter of the stress response in bacteria, transmits stress signals from the environment has remained elusive. The stressosome consists of multiple copies of three ...How the stressosome, the epicenter of the stress response in bacteria, transmits stress signals from the environment has remained elusive. The stressosome consists of multiple copies of three proteins RsbR, RsbS and RsbT, a kinase that is important for its activation. Using cryo-electron microscopy, we determined the atomic organization of the Listeria monocytogenes stressosome at 3.38 Å resolution. RsbR and RsbS are organized in a 60-protomers truncated icosahedron. A key phosphorylation site on RsbR (T209) is partially hidden by an RsbR flexible loop, whose "open" or "closed" position could modulate stressosome activity. Interaction between three glutamic acids in the N-terminal domain of RsbR and the membrane-bound mini-protein Prli42 is essential for Listeria survival to stress. Together, our data provide the atomic model of the stressosome core and highlight a loop important for stressosome activation, paving the way towards elucidating the mechanism of signal transduction by the stressosome in bacteria.
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_4510.map.gz / Format: CCP4 / Size: 262.9 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Listeria Stressosome
|Entire||Name: Listeria Stressosome / Details: The subcomponents are, RsbR, RsbR and he RsbT. / Number of components: 2|
-Component #1: protein, Listeria Stressosome
|Protein||Name: Listeria Stressosome / Details: The subcomponents are, RsbR, RsbR and he RsbT. / Recombinant expression: No|
|Mass||Experimental: 1.8 MDa|
|Source||Species: Listeria monocytogenes (bacteria) / Strain: EGD-e|
|Source (engineered)||Expression System: Escherichia coli (E. coli) / Strain: Bl21|
-Component #2: protein, RsbR
|Protein||Name: RsbR / Recombinant expression: No|
|Source||Species: Listeria monocytogenes EGD-E (bacteria)|
|Source (engineered)||Expression System: Escherichia coli (E. coli)|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 0.02 mg/mL / pH: 8.5|
|Vitrification||Cryogen name: OTHER|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: OTHER / Accelerating voltage: 300 kV / Electron dose: 2.25 e/Å2 / Illumination mode: OTHER|
|Lens||Imaging mode: OTHER|
|Specimen Holder||Model: FEI TITAN KRIOS AUTOGRID HOLDER|
|Camera||Detector: GATAN K2 QUANTUM (4k x 4k)|
|Processing||Method: single particle reconstruction / Number of projections: 78000|
|3D reconstruction||Software: RELION / Resolution: 3.37 Å / Resolution method: FSC 0.143 CUT-OFF|
|FSC plot (resolution estimation)|
-Atomic model buiding
|Modeling #1||Refinement space: REAL|
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