+
Open data
-
Basic information
Entry | Database: EMDB / ID: EMD-1757 | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
Title | The Structure of TubZ filaments | |||||||||
![]() | Double helical filaments of TubZ (pBT156) (Uniprot Q8KNP3) from Bacillus thuringiensis serovar israelensis (ATCC 35646) - negatively stained | |||||||||
![]() |
| |||||||||
![]() | Cytoskeletal / DNA segregation / FtsZ / FtsZ-like / pBtoxis / pBT156 / plasmid partitioning / RepX / tubulin / tubulin-like / TubZ | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | helical reconstruction / negative staining / Resolution: 35.0 Å | |||||||||
![]() | Aylett CHS / Amos LA / Lowe J | |||||||||
![]() | ![]() Title: Filament structure of bacterial tubulin homologue TubZ. Authors: Christopher H S Aylett / Qing Wang / Katharine A Michie / Linda A Amos / Jan Löwe / ![]() Abstract: Low copy number plasmids often depend on accurate partitioning systems for their continued survival. Generally, such systems consist of a centromere-like region of DNA, a DNA-binding adaptor, and a ...Low copy number plasmids often depend on accurate partitioning systems for their continued survival. Generally, such systems consist of a centromere-like region of DNA, a DNA-binding adaptor, and a polymerizing cytomotive filament. Together these components drive newly replicated plasmids to opposite ends of the dividing cell. The Bacillus thuringiensis plasmid pBToxis relies on a filament of the tubulin/FtsZ-like protein TubZ for its segregation. By combining crystallography and electron microscopy, we have determined the structure of this filament. We explain how GTP hydrolysis weakens the subunit-subunit contact and also shed light on the partitioning of the plasmid-adaptor complex. The double helical superstructure of TubZ filaments is unusual for tubulin-like proteins. Filaments of ParM, the actin-like partitioning protein, are also double helical. We suggest that convergent evolution shapes these different types of cytomotive filaments toward a general mechanism for plasmid separation. | |||||||||
History |
|
-
Structure visualization
Movie |
![]() |
---|---|
Structure viewer | EM map: ![]() ![]() ![]() |
Supplemental images |
-
Downloads & links
-EMDB archive
Map data | ![]() | 67.8 KB | ![]() | |
---|---|---|---|---|
Header (meta data) | ![]() ![]() | 8.4 KB 8.4 KB | Display Display | ![]() |
Images | ![]() | 341.1 KB | ||
Archive directory | ![]() ![]() | HTTPS FTP |
-Validation report
Summary document | ![]() | 203.7 KB | Display | ![]() |
---|---|---|---|---|
Full document | ![]() | 202.8 KB | Display | |
Data in XML | ![]() | 4 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
-
Links
EMDB pages | ![]() ![]() |
---|
-
Map
File | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Annotation | Double helical filaments of TubZ (pBT156) (Uniprot Q8KNP3) from Bacillus thuringiensis serovar israelensis (ATCC 35646) - negatively stained | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Voxel size | X=Y=Z: 4 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Density |
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Details | EMDB XML:
CCP4 map header:
|
-Supplemental data
-
Sample components
-Entire : Full length TubZ
Entire | Name: Full length TubZ |
---|---|
Components |
|
-Supramolecule #1000: Full length TubZ
Supramolecule | Name: Full length TubZ / type: sample / ID: 1000 / Details: Negatively stained / Oligomeric state: Double filament / Number unique components: 1 |
---|
-Macromolecule #1: Cytomotive filament
Macromolecule | Name: Cytomotive filament / type: protein_or_peptide / ID: 1 / Name.synonym: Cytomotive filament / Oligomeric state: Dimer / Recombinant expression: Yes |
---|---|
Source (natural) | Organism: ![]() ![]() |
Recombinant expression | Organism: ![]() ![]() |
-Experimental details
-Structure determination
Method | negative staining |
---|---|
![]() | helical reconstruction |
Aggregation state | filament |
-
Sample preparation
Concentration | 0.1 mg/mL |
---|---|
Buffer | pH: 7.5 / Details: 50 mM NaHEPES 7.5 150 mM KCl 5 mM MgCl2 1 mM GTPyS |
Staining | Type: NEGATIVE / Details: 1% Uranyl Acetate |
Grid | Details: CuRh 300 mesh |
Vitrification | Cryogen name: NONE / Instrument: OTHER |
-
Electron microscopy
Microscope | FEI TECNAI 12 |
---|---|
Temperature | Average: 293 K |
Image recording | Category: CCD / Film or detector model: KODAK SO-163 FILM |
Electron beam | Acceleration voltage: 120 kV / Electron source: TUNGSTEN HAIRPIN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal magnification: 69000 |
Sample stage | Specimen holder: Eucentric / Specimen holder model: SIDE ENTRY, EUCENTRIC |
-
Image processing
Final reconstruction | Applied symmetry - Helical parameters - Axial symmetry: C2 (2 fold cyclic) Algorithm: OTHER / Resolution.type: BY AUTHOR / Resolution: 35.0 Å / Resolution method: OTHER / Software - Name: MRC Details: Final maps were calculated from five averaged datasets |
---|