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- EMDB-18004: Cryo electron tomogram of Caulobacter crescentus - Delta-bla -

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Basic information

Entry
Database: EMDB / ID: EMD-18004
TitleCryo electron tomogram of Caulobacter crescentus - Delta-bla
Map data
Sample
  • Cell: Caulobacter crescentus Delta-bla mutant
KeywordsAdaptive multi-drug resistance / Efflux pump / Co-chaperone / Beta-lactam / TipR / DjlA / MDR / Caulobacter crescentus / ANTIBIOTIC
Biological speciesCaulobacter vibrioides CB15 (bacteria)
Methodelectron tomography / cryo EM
AuthorsCostafrolaz J / Panis G / Casu B / Ardissone S / Degeorges L / Pilhofer M / Viollier PH
Funding support Switzerland, European Union, 2 items
OrganizationGrant numberCountry
Swiss National Science Foundation31003A_179255 Switzerland
European Research Council (ERC)679209European Union
CitationJournal: PLoS Biol / Year: 2023
Title: Adaptive β-lactam resistance from an inducible efflux pump that is post-translationally regulated by the DjlA co-chaperone.
Authors: Jordan Costafrolaz / Gaël Panis / Bastien Casu / Silvia Ardissone / Laurence Degeorges / Martin Pilhofer / Patrick H Viollier /
Abstract: The acquisition of multidrug resistance (MDR) determinants jeopardizes treatment of bacterial infections with antibiotics. The tripartite efflux pump AcrAB-NodT confers adaptive MDR in the polarized ...The acquisition of multidrug resistance (MDR) determinants jeopardizes treatment of bacterial infections with antibiotics. The tripartite efflux pump AcrAB-NodT confers adaptive MDR in the polarized α-proteobacterium Caulobacter crescentus via transcriptional induction by first-generation quinolone antibiotics. We discovered that overexpression of AcrAB-NodT by mutation or exogenous inducers confers resistance to cephalosporin and penicillin (β-lactam) antibiotics. Combining 2-step mutagenesis-sequencing (Mut-Seq) and cephalosporin-resistant point mutants, we dissected how TipR uses a common operator of the divergent tipR and acrAB-nodT promoter for adaptive and/or potentiated AcrAB-NodT-directed efflux. Chemical screening identified diverse compounds that interfere with DNA binding by TipR or induce its dependent proteolytic turnover. We found that long-term induction of AcrAB-NodT deforms the envelope and that homeostatic control by TipR includes co-induction of the DnaJ-like co-chaperone DjlA, boosting pump assembly and/or capacity in anticipation of envelope stress. Thus, the adaptive MDR regulatory circuitry reconciles drug efflux with co-chaperone function for trans-envelope assemblies and maintenance.
History
DepositionJul 24, 2023-
Header (metadata) releaseNov 15, 2023-
Map releaseNov 15, 2023-
UpdateMay 29, 2024-
Current statusMay 29, 2024Processing site: PDBe / Status: Released

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Structure visualization

Supplemental images

emd_18004.png

Downloads & links

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Map

FileDownload / File: emd_18004.map.gz / Format: CCP4 / Size: 322.9 MB / Type: IMAGE STORED AS SIGNED BYTE
Voxel sizeX=Y=Z: 11 Å
Density
Minimum - Maximum-128.0 - 127.0
Average (Standard dev.)11.118971999999999 (±17.700437999999998)
SymmetrySpace group: 1
Details

EMDB XML:

Map geometry
Axis orderXYZ
Origin01190
Dimensions928960380
Spacing960928380
CellA: 10560.0 Å / B: 10208.0 Å / C: 4180.0 Å
α=β=γ: 90.0 °

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Supplemental data

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Sample components

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Entire : Caulobacter crescentus Delta-bla mutant

EntireName: Caulobacter crescentus Delta-bla mutant
Components
  • Cell: Caulobacter crescentus Delta-bla mutant

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Supramolecule #1: Caulobacter crescentus Delta-bla mutant

SupramoleculeName: Caulobacter crescentus Delta-bla mutant / type: cell / ID: 1 / Parent: 0
Source (natural)Organism: Caulobacter vibrioides CB15 (bacteria)

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Experimental details

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Structure determination

Methodcryo EM
Processingelectron tomography
Aggregation statecell

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Sample preparation

BufferpH: 7
VitrificationCryogen name: ETHANE-PROPANE / Instrument: FEI VITROBOT MARK IV
SectioningOther: NO SECTIONING
Fiducial markerManufacturer: cytodiagnostics / Diameter: 10 nm

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Electron microscopy

MicroscopeFEI TITAN KRIOS
Electron beamAcceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN
Electron opticsIllumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELDBright-field microscopy / Nominal defocus max: 8.0 µm / Nominal defocus min: 8.0 µm
Image recordingFilm or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: COUNTING / Number real images: 61 / Average electron dose: 2.1 e/Å2 / Details: 61 tilt series with around 130 accumulated dose.
Experimental equipment
Model: Titan Krios / Image courtesy: FEI Company

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Image processing

Final reconstructionSoftware - Name: IMOD / Number images used: 61

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