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Open data
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Basic information
Entry | Database: PDB / ID: 8aaj | |||||||||
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Title | Crystal structure of the Pyrococcus abyssi RPA (apo form) | |||||||||
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![]() | DNA BINDING PROTEIN / Replication protein A / ssDNA-Binding protein | |||||||||
Function / homology | ![]() response to ionizing radiation / double-strand break repair via homologous recombination / nucleic acid binding / chromosome, telomeric region / DNA binding / metal ion binding Similarity search - Function | |||||||||
Biological species | ![]() ![]() | |||||||||
Method | ![]() ![]() ![]() | |||||||||
![]() | Legrand, P. / Madru, C. / Sauguet, L. | |||||||||
Funding support | ![]()
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![]() | ![]() Title: DNA-binding mechanism and evolution of replication protein A. Authors: Clément Madru / Markel Martínez-Carranza / Sébastien Laurent / Alessandra C Alberti / Maelenn Chevreuil / Bertrand Raynal / Ahmed Haouz / Rémy A Le Meur / Marc Delarue / Ghislaine ...Authors: Clément Madru / Markel Martínez-Carranza / Sébastien Laurent / Alessandra C Alberti / Maelenn Chevreuil / Bertrand Raynal / Ahmed Haouz / Rémy A Le Meur / Marc Delarue / Ghislaine Henneke / Didier Flament / Mart Krupovic / Pierre Legrand / Ludovic Sauguet / ![]() Abstract: Replication Protein A (RPA) is a heterotrimeric single stranded DNA-binding protein with essential roles in DNA replication, recombination and repair. Little is known about the structure of RPA in ...Replication Protein A (RPA) is a heterotrimeric single stranded DNA-binding protein with essential roles in DNA replication, recombination and repair. Little is known about the structure of RPA in Archaea, the third domain of life. By using an integrative structural, biochemical and biophysical approach, we extensively characterize RPA from Pyrococcus abyssi in the presence and absence of DNA. The obtained X-ray and cryo-EM structures reveal that the trimerization core and interactions promoting RPA clustering on ssDNA are shared between archaea and eukaryotes. However, we also identified a helical domain named AROD (Acidic Rpa1 OB-binding Domain), and showed that, in Archaea, RPA forms an unanticipated tetrameric supercomplex in the absence of DNA. The four RPA molecules clustered within the tetramer could efficiently coat and protect stretches of ssDNA created by the advancing replisome. Finally, our results provide insights into the evolution of this primordial replication factor in eukaryotes. | |||||||||
History |
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Structure visualization
Structure viewer | Molecule: ![]() ![]() |
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Downloads & links
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Download
PDBx/mmCIF format | ![]() | 283.6 KB | Display | ![]() |
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PDB format | ![]() | 232.8 KB | Display | ![]() |
PDBx/mmJSON format | ![]() | Tree view | ![]() | |
Others | ![]() |
-Validation report
Summary document | ![]() | 513.3 KB | Display | ![]() |
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Full document | ![]() | 517.5 KB | Display | |
Data in XML | ![]() | 23.8 KB | Display | |
Data in CIF | ![]() | 32 KB | Display | |
Arichive directory | ![]() ![]() | HTTPS FTP |
-Related structure data
Related structure data | ![]() 8aa9C ![]() 8aasC ![]() 8c5yC ![]() 8c5zC ![]() 8oejC ![]() 8oelC C: citing same article ( |
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Similar structure data | Similarity search - Function & homology ![]() |
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Links
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Assembly
Deposited unit | ![]()
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Unit cell |
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Components
#1: Protein | Mass: 41008.965 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: Mutation T2S from reference sequence GenBank: CCE69663.1 to improve expression Source: (gene. exp.) ![]() ![]() ![]() ![]() | ||
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#2: Protein | Mass: 31376.152 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Details: Mutation of SER after first M from reference sequence NCBI Reference Sequence: WP_048146526.1 to improve expression Source: (gene. exp.) ![]() ![]() ![]() ![]() | ||
#3: Protein | Mass: 14008.925 Da / Num. of mol.: 1 Source method: isolated from a genetically manipulated source Source: (gene. exp.) ![]() ![]() ![]() ![]() | ||
#4: Chemical | ChemComp-ZN / | ||
#5: Chemical | ChemComp-SO4 / Has ligand of interest | Y | |
-Experimental details
-Experiment
Experiment | Method: ![]() |
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Sample preparation
Crystal | Density Matthews: 5.78 Å3/Da / Density % sol: 78.71 % |
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Crystal grow | Temperature: 291 K / Method: vapor diffusion, hanging drop / pH: 8 / Details: 0.1M Tris pH 8.5 1.2M ammonium sulfate |
-Data collection
Diffraction | Mean temperature: 100 K / Serial crystal experiment: N |
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Diffraction source | Source: ![]() ![]() ![]() |
Detector | Type: DECTRIS EIGER X 16M / Detector: PIXEL / Date: Mar 28, 2021 / Details: KB Mirrors |
Radiation | Monochromator: Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray |
Radiation wavelength | Wavelength: 0.978565 Å / Relative weight: 1 |
Reflection | Resolution: 3.7→49.35 Å / Num. obs: 13191 / % possible obs: 60.1 % / Redundancy: 27.6 % / Biso Wilson estimate: 166.58 Å2 / CC1/2: 1 / Rmerge(I) obs: 0.082 / Rpim(I) all: 0.016 / Rrim(I) all: 0.083 / Net I/σ(I): 21.4 |
Reflection shell | Resolution: 3.7→3.8 Å / Redundancy: 26.8 % / Rmerge(I) obs: 4.306 / Mean I/σ(I) obs: 1 / Num. unique obs: 217 / CC1/2: 0.542 / Rpim(I) all: 0.844 / Rrim(I) all: 4.39 / % possible all: 13.3 |
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Processing
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Refinement | Method to determine structure: ![]()
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Displacement parameters | Biso mean: 235.87 Å2
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Refine analyze | Luzzati coordinate error obs: 0.64 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Refinement step | Cycle: LAST / Resolution: 3.7→49.35 Å
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Refine LS restraints |
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LS refinement shell | Resolution: 3.7→3.81 Å
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Refinement TLS params. | Refine-ID: X-RAY DIFFRACTION
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Refinement TLS group |
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