[English] 日本語
Yorodumi- PDB-8bl4: Cryo-EM structure of a contractile injection system in Streptomyc... -
+Open data
-Basic information
Entry | Database: PDB / ID: 8bl4 | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of a contractile injection system in Streptomyces coelicolor, the sheath-tube module in extended state. | ||||||||||||
Components |
| ||||||||||||
Keywords | STRUCTURAL PROTEIN / Contractile injection system | ||||||||||||
Function / homology | Function and homology information | ||||||||||||
Biological species | Streptomyces coelicolor A3 | ||||||||||||
Method | ELECTRON MICROSCOPY / helical reconstruction / cryo EM / Resolution: 3.9 Å | ||||||||||||
Authors | Casu, B. / Sallmen, J.W. / Schlimpert, S. / Pilhofer, M. | ||||||||||||
Funding support | Switzerland, European Union, United Kingdom, 3items
| ||||||||||||
Citation | Journal: Nat Microbiol / Year: 2023 Title: Cytoplasmic contractile injection systems mediate cell death in Streptomyces. Authors: Bastien Casu / Joseph W Sallmen / Susan Schlimpert / Martin Pilhofer / Abstract: Contractile injection systems (CIS) are bacteriophage tail-like structures that mediate bacterial cell-cell interactions. While CIS are highly abundant across diverse bacterial phyla, representative ...Contractile injection systems (CIS) are bacteriophage tail-like structures that mediate bacterial cell-cell interactions. While CIS are highly abundant across diverse bacterial phyla, representative gene clusters in Gram-positive organisms remain poorly studied. Here we characterize a CIS in the Gram-positive multicellular model organism Streptomyces coelicolor and show that, in contrast to most other CIS, S. coelicolor CIS (CIS) mediate cell death in response to stress and impact cellular development. CIS are expressed in the cytoplasm of vegetative hyphae and are not released into the medium. Our cryo-electron microscopy structure enabled the engineering of non-contractile and fluorescently tagged CIS assemblies. Cryo-electron tomography showed that CIS contraction is linked to reduced cellular integrity. Fluorescence light microscopy furthermore revealed that functional CIS mediate cell death upon encountering different types of stress. The absence of functional CIS had an impact on hyphal differentiation and secondary metabolite production. Finally, we identified three putative effector proteins, which when absent, phenocopied other CIS mutants. Our results provide new functional insights into CIS in Gram-positive organisms and a framework for studying novel intracellular roles, including regulated cell death and life-cycle progression in multicellular bacteria. | ||||||||||||
History |
|
-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
---|
-Downloads & links
-Download
PDBx/mmCIF format | 8bl4.cif.gz | 2.1 MB | Display | PDBx/mmCIF format |
---|---|---|---|---|
PDB format | pdb8bl4.ent.gz | 1.7 MB | Display | PDB format |
PDBx/mmJSON format | 8bl4.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 8bl4_validation.pdf.gz | 1.7 MB | Display | wwPDB validaton report |
---|---|---|---|---|
Full document | 8bl4_full_validation.pdf.gz | 1.7 MB | Display | |
Data in XML | 8bl4_validation.xml.gz | 285.6 KB | Display | |
Data in CIF | 8bl4_validation.cif.gz | 449.6 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/bl/8bl4 ftp://data.pdbj.org/pub/pdb/validation_reports/bl/8bl4 | HTTPS FTP |
-Related structure data
Related structure data | 16101MC 8bkyC M: map data used to model this data C: citing same article (ref.) |
---|---|
Similar structure data | Similarity search - Function & homologyF&H Search |
-Links
-Assembly
Deposited unit |
|
---|---|
1 |
|
-Components
#1: Protein | Mass: 57465.113 Da / Num. of mol.: 24 / Mutation: Insertion 26-IEGVG / Source method: isolated from a natural source / Source: (natural) Streptomyces coelicolor A3(2) (bacteria) / References: UniProt: Q9L0N8 #2: Protein | Mass: 16493.668 Da / Num. of mol.: 24 / Source method: isolated from a natural source / Source: (natural) Streptomyces coelicolor A3(2) (bacteria) / References: UniProt: Q9L0N9 |
---|
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
---|---|
EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: helical reconstruction |
-Sample preparation
Component | Name: The sheath-tube module of a contractile injection system in Streptomyces coelicolor Type: COMPLEX / Entity ID: all / Source: NATURAL |
---|---|
Source (natural) | Organism: Streptomyces coelicolor A3(2) (bacteria) |
Buffer solution | pH: 8 |
Specimen | Embedding applied: NO / Shadowing applied: NO / Staining applied: NO / Vitrification applied: YES |
Vitrification | Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE-PROPANE |
-Electron microscopy imaging
Experimental equipment | 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 FIELD / Nominal defocus max: 3500 nm / Nominal defocus min: 1500 nm |
Image recording | Electron dose: 60 e/Å2 / Film or detector model: GATAN K2 SUMMIT (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
---|---|
Helical symmerty | Angular rotation/subunit: 23.1 ° / Axial rise/subunit: 38.5 Å / Axial symmetry: C6 |
3D reconstruction | Resolution: 3.9 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 18822 / Symmetry type: HELICAL |