|Entry||Database: PDB / ID: 6yg8|
|Title||Cryo-EM structure of a BcsB pentamer in the context of an assembled Bcs macrocomplex|
|Components||Bacterial cellulose secretion regulator BcsB|
|Keywords||SIGNALING PROTEIN / Bacterial biofilms / Bacterial cellulose / Bacterial secretion system / regulator protein|
|Function / homology||Cellulose synthase, subunit B / Galactose-binding-like domain superfamily / Cellulose synthase BcsB, bacterial / cellulose biosynthetic process / UDP-glucose metabolic process / integral component of membrane / plasma membrane / Cyclic di-GMP-binding protein|
Function and homology information
|Biological species||Escherichia coli (E. coli)|
|Method||ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3 Å|
|Authors||Zouhir, S. / Krasteva, P.V.|
|Funding support||5items |
|Citation||Journal: Sci Adv / Year: 2021|
Title: Architecture and regulation of an enterobacterial cellulose secretion system.
Authors: Wiem Abidi / Samira Zouhir / Meryem Caleechurn / Stéphane Roche / Petya Violinova Krasteva /
Abstract: Many free-living and pathogenic enterobacteria secrete biofilm-promoting cellulose using a multicomponent, envelope-embedded Bcs secretion system under the control of intracellular second messenger c- ...Many free-living and pathogenic enterobacteria secrete biofilm-promoting cellulose using a multicomponent, envelope-embedded Bcs secretion system under the control of intracellular second messenger c-di-GMP. The molecular understanding of system assembly and cellulose secretion has been largely limited to the crystallographic studies of a distantly homologous BcsAB synthase tandem and a low-resolution reconstruction of an assembled macrocomplex that encompasses most of the inner membrane and cytosolic subunits and features an atypical layered architecture. Here, we present cryo-EM structures of the assembled Bcs macrocomplex, as well as multiple crystallographic snapshots of regulatory Bcs subcomplexes. The structural and functional data uncover the mechanism of asymmetric secretion system assembly and periplasmic crown polymerization and reveal unexpected subunit stoichiometry, multisite c-di-GMP recognition, and ATP-dependent regulation.
SummaryFull reportAbout validation report
|Structure viewer||Molecule: |
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A: Bacterial cellulose secretion regulator BcsB
B: Bacterial cellulose secretion regulator BcsB
C: Bacterial cellulose secretion regulator BcsB
D: Bacterial cellulose secretion regulator BcsB
E: Bacterial cellulose secretion regulator BcsB
Mass: 86184.383 Da / Num. of mol.: 5
Source method: isolated from a genetically manipulated source
Details: Bacterial cellulose synthesis subunit B / Source: (gene. exp.) Escherichia coli (E. coli) / Strain: 1094
Gene: bcsB, A8C65_00280, AC789_1c39010, ACU57_05345, AM464_10380, BHS81_21120, BK292_24560, BON72_13325, BON75_10020, BON76_21165, BON94_21155, BZL31_14275, C5P01_24165, C9162_26260, CI641_010915, ...Gene: bcsB, A8C65_00280, AC789_1c39010, ACU57_05345, AM464_10380, BHS81_21120, BK292_24560, BON72_13325, BON75_10020, BON76_21165, BON94_21155, BZL31_14275, C5P01_24165, C9162_26260, CI641_010915, CWS33_18410, D3O91_11715, D9I18_15765, DQF57_09480, E0J34_09745, E0K84_22715, E2127_16410, E2128_18000, E2129_18135, E2134_17800, E5P22_13645, ELV08_08610, F1E03_18495, F1E19_10130, FRV13_18850
Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: A0A061KLG7
|Experiment||Method: ELECTRON MICROSCOPY|
|EM experiment||Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction|
|Molecular weight||Value: 0.734 MDa / Experimental value: NO|
|Buffer solution||pH: 8|
|Specimen||Conc.: 0.8 mg/ml / Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Humidity: 100 % / Chamber temperature: 277 K|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Microscopy||Model: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Illumination mode: FLOOD BEAM|
|Electron lens||Mode: BRIGHT FIELDBright-field microscopy / Nominal magnification: 130000 X / Nominal defocus max: 2750 nm / Nominal defocus min: 750 nm / Cs: 2.7 mm / C2 aperture diameter: 100 µm|
|Image recording||Electron dose: 1.2 e/Å2 / Detector mode: COUNTING / Film or detector model: GATAN K2 SUMMIT (4k x 4k)|
|EM imaging optics||Energyfilter name: GIF Quantum LS|
|Software||Name: PHENIX / Version: 1.16_3549: / Classification: refinement|
|Image processing||Details: A total of 9,129 movies recorded on the CM01 Titan Krios transmission electron microscope (Thermo Fisher Scientific) at the ESRF Grenoble operated at 300 kV and equipped with a Gatan K2 ...Details: A total of 9,129 movies recorded on the CM01 Titan Krios transmission electron microscope (Thermo Fisher Scientific) at the ESRF Grenoble operated at 300 kV and equipped with a Gatan K2 Summit direct electron detector and a GIF Quantum LS imaging filter|
|CTF correction||Details: Gctf through the cryoSPARC v2 interface. / Type: PHASE FLIPPING AND AMPLITUDE CORRECTION|
|Symmetry||Point symmetry: C1 (asymmetric)|
|3D reconstruction||Resolution: 3 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 576455 / Num. of class averages: 1 / Symmetry type: POINT|
|Atomic model building||Space: REAL|
|Refine LS restraints|
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