[English] 日本語
Yorodumi- PDB-7vpd: Cryo-EM structure of Streptomyces coelicolor RNAP-promoter open c... -
+Open data
-Basic information
Entry | Database: PDB / ID: 7vpd | ||||||
---|---|---|---|---|---|---|---|
Title | Cryo-EM structure of Streptomyces coelicolor RNAP-promoter open complex with one Zur dimers | ||||||
Components |
| ||||||
Keywords | DNA BINDING PROTEIN | ||||||
Function / homology | Function and homology information regulation of secondary metabolite biosynthetic process / DNA-binding transcription repressor activity / sigma factor activity / DNA-directed RNA polymerase complex / protein-DNA complex / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / protein dimerization activity ...regulation of secondary metabolite biosynthetic process / DNA-binding transcription repressor activity / sigma factor activity / DNA-directed RNA polymerase complex / protein-DNA complex / DNA-templated transcription initiation / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / DNA-directed RNA polymerase / protein dimerization activity / transcription cis-regulatory region binding / DNA-binding transcription factor activity / negative regulation of DNA-templated transcription / DNA-templated transcription / magnesium ion binding / DNA binding / zinc ion binding / cytosol / cytoplasm Similarity search - Function | ||||||
Biological species | Streptomyces coelicolor A3 | ||||||
Method | ELECTRON MICROSCOPY / single particle reconstruction / cryo EM / Resolution: 3.77 Å | ||||||
Authors | Yang, X. / Zheng, J. | ||||||
Funding support | China, 1items
| ||||||
Citation | Journal: Nucleic Acids Res / Year: 2022 Title: Structural basis of Streptomyces transcription activation by zinc uptake regulator. Authors: Xu Yang / Yiqun Wang / Guiyang Liu / Zixin Deng / Shuangjun Lin / Jianting Zheng / Abstract: Streptomyces coelicolor (Sc) is a model organism of actinobacteria to study morphological differentiation and production of bioactive metabolites. Sc zinc uptake regulator (Zur) affects both ...Streptomyces coelicolor (Sc) is a model organism of actinobacteria to study morphological differentiation and production of bioactive metabolites. Sc zinc uptake regulator (Zur) affects both processes by controlling zinc homeostasis. It activates transcription by binding to palindromic Zur-box sequences upstream of -35 elements. Here we deciphered the molecular mechanism by which ScZur interacts with promoter DNA and Sc RNA polymerase (RNAP) by cryo-EM structures and biochemical assays. The ScZur-DNA structures reveal a sequential and cooperative binding of three ScZur dimers surrounding a Zur-box spaced 8 nt upstream from a -35 element. The ScRNAPσHrdB-Zur-DNA structures define protein-protein and protein-DNA interactions involved in the principal housekeeping σHrdB-dependent transcription initiation from a noncanonical promoter with a -10 element lacking the critical adenine residue at position -11 and a TTGCCC -35 element deviating from the canonical TTGACA motif. ScZur interacts with the C-terminal domain of ScRNAP α subunit (αCTD) in a complex structure trapped in an active conformation. Key ScZur-αCTD interfacial residues accounting for ScZur-dependent transcription activation were confirmed by mutational studies. Together, our structural and biochemical results provide a comprehensive model for transcription activation of Zur family regulators. | ||||||
History |
|
-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
---|
-Downloads & links
-Download
PDBx/mmCIF format | 7vpd.cif.gz | 709 KB | Display | PDBx/mmCIF format |
---|---|---|---|---|
PDB format | pdb7vpd.ent.gz | 559.7 KB | Display | PDB format |
PDBx/mmJSON format | 7vpd.json.gz | Tree view | PDBx/mmJSON format | |
Others | Other downloads |
-Validation report
Summary document | 7vpd_validation.pdf.gz | 1.4 MB | Display | wwPDB validaton report |
---|---|---|---|---|
Full document | 7vpd_full_validation.pdf.gz | 1.4 MB | Display | |
Data in XML | 7vpd_validation.xml.gz | 102.2 KB | Display | |
Data in CIF | 7vpd_validation.cif.gz | 160.9 KB | Display | |
Arichive directory | https://data.pdbj.org/pub/pdb/validation_reports/vp/7vpd ftp://data.pdbj.org/pub/pdb/validation_reports/vp/7vpd | HTTPS FTP |
-Related structure data
Related structure data | 32063MC 7vo0C 7vo9C 7vpzC 7x74C 7x75C 7x76C 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
-DNA-directed RNA polymerase subunit ... , 4 types, 6 molecules ABSCDE
#1: Protein | Mass: 36734.641 Da / Num. of mol.: 3 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Streptomyces coelicolor A3(2) (bacteria) Gene: rpoA, GTW64_13255 / Production host: Escherichia coli BL21(DE3) (bacteria) References: UniProt: A0A6G2M9E1, DNA-directed RNA polymerase #2: Protein | | Mass: 128644.945 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Streptomyces coelicolor A3(2) (bacteria) Strain: ATCC BAA-471 / A3(2) / M145 / Gene: rpoB, SCO4654, SCD82.26 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9L0L0, DNA-directed RNA polymerase #3: Protein | | Mass: 145912.219 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Streptomyces coelicolor A3(2) (bacteria) Strain: ATCC BAA-471 / A3(2) / M145 / Gene: rpoC, SCO4655, SCD40A.01, SCD82.27 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q8CJT1, DNA-directed RNA polymerase #4: Protein | | Mass: 9716.941 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Streptomyces coelicolor A3(2) (bacteria) Strain: ATCC BAA-471 / A3(2) / M145 / Gene: rpoZ, SCO1478, SC9C5.02c / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9KXS1, DNA-directed RNA polymerase |
---|
-Protein , 2 types, 3 molecules FMN
#5: Protein | Mass: 58288.078 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Streptomyces coelicolor A3(2) (bacteria) Gene: sigA, SLI_6088 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: A0A7U9DYK1 |
---|---|
#6: Protein | Mass: 16904.840 Da / Num. of mol.: 2 Source method: isolated from a genetically manipulated source Source: (gene. exp.) Streptomyces coelicolor A3(2) (bacteria) Strain: ATCC BAA-471 / A3(2) / M145 / Gene: SCO2508 / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9L2H5 |
-DNA chain , 2 types, 2 molecules OP
#7: DNA chain | Mass: 25808.424 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Streptomyces coelicolor A3(2) (bacteria) / References: GenBank: 24418971 |
---|---|
#8: DNA chain | Mass: 26017.555 Da / Num. of mol.: 1 / Source method: obtained synthetically / Source: (synth.) Streptomyces coelicolor A3(2) (bacteria) |
-Non-polymers , 2 types, 9 molecules
#9: Chemical | ChemComp-MG / |
---|---|
#10: Chemical | ChemComp-ZN / |
-Details
Has ligand of interest | Y |
---|
-Experimental details
-Experiment
Experiment | Method: ELECTRON MICROSCOPY |
---|---|
EM experiment | Aggregation state: PARTICLE / 3D reconstruction method: single particle reconstruction |
-Sample preparation
Component | Name: Cryo-EM structure of Streptomyces coelicolor RNAP-promoter open complex with one Zur dimers Type: COMPLEX / Entity ID: #1-#8 / Source: RECOMBINANT | ||||||||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Molecular weight | Value: 0.56 MDa / Experimental value: YES | ||||||||||||||||||||||||||||||
Source (natural) | Organism: Streptomyces coelicolor A3(2) (bacteria) | ||||||||||||||||||||||||||||||
Source (recombinant) | Organism: Escherichia coli BL21(DE3) (bacteria) | ||||||||||||||||||||||||||||||
Buffer solution | pH: 8 | ||||||||||||||||||||||||||||||
Buffer component |
| ||||||||||||||||||||||||||||||
Specimen | Conc.: 1 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: 289 K |
-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: 2000 nm / Nominal defocus min: 1000 nm |
Image recording | Electron dose: 40 e/Å2 / Film or detector model: GATAN K2 QUANTUM (4k x 4k) |
-Processing
CTF correction | Type: PHASE FLIPPING AND AMPLITUDE CORRECTION |
---|---|
3D reconstruction | Resolution: 3.77 Å / Resolution method: FSC 0.143 CUT-OFF / Num. of particles: 19427 / Symmetry type: POINT |