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- PDB-7f90: Crystal structure of SARS auxiliary protein in complex with human... -

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Basic information

Entry
Database: PDB / ID: 7f90
TitleCrystal structure of SARS auxiliary protein in complex with human nuclear protein
Components
  • Nuclear pore complex protein Nup98-Nup96Nuclear pore
  • ORF6 protein
  • mRNA export factor
KeywordsVIRAL PROTEIN / immune system / auxiliary protein
Function / homology
Function and homology information


transcription-dependent tethering of RNA polymerase II gene DNA at nuclear periphery / telomere tethering at nuclear periphery / nuclear pore organization / nuclear pore outer ring / nuclear pore complex assembly / post-transcriptional tethering of RNA polymerase II gene DNA at nuclear periphery / nuclear pore cytoplasmic filaments / Nuclear Pore Complex (NPC) Disassembly / nuclear inclusion body / nuclear pore nuclear basket ...transcription-dependent tethering of RNA polymerase II gene DNA at nuclear periphery / telomere tethering at nuclear periphery / nuclear pore organization / nuclear pore outer ring / nuclear pore complex assembly / post-transcriptional tethering of RNA polymerase II gene DNA at nuclear periphery / nuclear pore cytoplasmic filaments / Nuclear Pore Complex (NPC) Disassembly / nuclear inclusion body / nuclear pore nuclear basket / Transport of Ribonucleoproteins into the Host Nucleus / Regulation of Glucokinase by Glucokinase Regulatory Protein / Defective TPR may confer susceptibility towards thyroid papillary carcinoma (TPC) / Transport of the SLBP independent Mature mRNA / Transport of the SLBP Dependant Mature mRNA / NS1 Mediated Effects on Host Pathways / SUMOylation of SUMOylation proteins / Transport of Mature mRNA Derived from an Intronless Transcript / structural constituent of nuclear pore / Rev-mediated nuclear export of HIV RNA / positive regulation of mRNA splicing, via spliceosome / SUMOylation of RNA binding proteins / Nuclear import of Rev protein / RNA export from nucleus / Transport of Mature mRNA derived from an Intron-Containing Transcript / NEP/NS2 Interacts with the Cellular Export Machinery / tRNA processing in the nucleus / Postmitotic nuclear pore complex (NPC) reformation / nucleocytoplasmic transport / Viral Messenger RNA Synthesis / nuclear localization sequence binding / SUMOylation of ubiquitinylation proteins / mitotic spindle pole / host cell Golgi membrane / Vpr-mediated nuclear import of PICs / SUMOylation of DNA replication proteins / cellular response to organic cyclic compound / Hydrolases; Acting on peptide bonds (peptidases); Serine endopeptidases / Regulation of HSF1-mediated heat shock response / mRNA transport / Amplification of signal from unattached kinetochores via a MAD2 inhibitory signal / mRNA export from nucleus / SUMOylation of DNA damage response and repair proteins / nuclear pore / Mitotic Prometaphase / EML4 and NUDC in mitotic spindle formation / symbiont-mediated suppression of host JAK-STAT cascade via inhibition of STAT1 activity / regulation of mitotic spindle organization / Resolution of Sister Chromatid Cohesion / serine-type peptidase activity / SUMOylation of chromatin organization proteins / nuclear periphery / HCMV Late Events / ubiquitin binding / RHO GTPases Activate Formins / promoter-specific chromatin binding / Transcriptional regulation by small RNAs / ISG15 antiviral mechanism / fibrillar center / HCMV Early Events / protein import into nucleus / Separation of Sister Chromatids / nuclear envelope / snRNP Assembly / microtubule binding / nuclear membrane / transcription coactivator activity / host cell endoplasmic reticulum membrane / nuclear body / cell cycle / ribonucleoprotein complex / cell division / mRNA binding / symbiont-mediated suppression of host type I interferon-mediated signaling pathway / SARS-CoV-2 activates/modulates innate and adaptive immune responses / proteolysis / RNA binding / nucleoplasm / membrane / nucleus / cytosol / cytoplasm
Similarity search - Function
Non-structural protein NS6, betacoronavirus / Betacoronavirus NS6 protein / Nup98, Gle2-binding sequence / Nuclear pore complex protein NUP96, C-terminal domain / Nuclear protein 96 / Nuclear pore complex protein Nup98-Nup96-like, autopeptidase S59 domain / Nuclear pore complex protein Nup98-Nup96-like, autopeptidase S59 domain superfamily / Nucleoporin autopeptidase / NUP C-terminal domain profile. / Nucleoporin peptidase S59-like ...Non-structural protein NS6, betacoronavirus / Betacoronavirus NS6 protein / Nup98, Gle2-binding sequence / Nuclear pore complex protein NUP96, C-terminal domain / Nuclear protein 96 / Nuclear pore complex protein Nup98-Nup96-like, autopeptidase S59 domain / Nuclear pore complex protein Nup98-Nup96-like, autopeptidase S59 domain superfamily / Nucleoporin autopeptidase / NUP C-terminal domain profile. / Nucleoporin peptidase S59-like / G-protein beta WD-40 repeat / WD40 repeat, conserved site / Trp-Asp (WD) repeats signature. / WD domain, G-beta repeat / WD40 repeats / WD40 repeat / Trp-Asp (WD) repeats profile. / Trp-Asp (WD) repeats circular profile. / WD40-repeat-containing domain superfamily / WD40/YVTN repeat-like-containing domain superfamily
Similarity search - Domain/homology
Nuclear pore complex protein Nup98-Nup96 / ORF6 protein / mRNA export factor RAE1
Similarity search - Component
Biological speciesSevere acute respiratory syndrome coronavirus
Homo sapiens (human)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.39 Å
AuthorsGao, X. / Cui, S.
CitationJournal: Nat Commun / Year: 2022
Title: Structural basis for Sarbecovirus ORF6 mediated blockage of nucleocytoplasmic transport
Authors: Gao, X. / Tian, H. / Zhu, K. / Li, Q. / Hao, W. / Wang, L. / Qin, B. / Deng, H. / Cui, S.
History
DepositionJul 3, 2021Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0Sep 14, 2022Provider: repository / Type: Initial release
Revision 1.1Nov 29, 2023Group: Data collection / Refinement description
Category: chem_comp_atom / chem_comp_bond / pdbx_initial_refinement_model

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
E: ORF6 protein
F: ORF6 protein
A: mRNA export factor
C: mRNA export factor
B: Nuclear pore complex protein Nup98-Nup96
D: Nuclear pore complex protein Nup98-Nup96


Theoretical massNumber of molelcules
Total (without water)492,6636
Polymers492,6636
Non-polymers00
Water3,405189
1
E: ORF6 protein
A: mRNA export factor
B: Nuclear pore complex protein Nup98-Nup96


Theoretical massNumber of molelcules
Total (without water)246,3313
Polymers246,3313
Non-polymers00
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3870 Å2
ΔGint-15 kcal/mol
Surface area16610 Å2
MethodPISA
2
F: ORF6 protein
C: mRNA export factor
D: Nuclear pore complex protein Nup98-Nup96


Theoretical massNumber of molelcules
Total (without water)246,3313
Polymers246,3313
Non-polymers00
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area4010 Å2
ΔGint-18 kcal/mol
Surface area16420 Å2
MethodPISA
Unit cell
Length a, b, c (Å)189.332, 86.794, 47.905
Angle α, β, γ (deg.)90.000, 92.090, 90.000
Int Tables number5
Space group name H-MC121

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Components

#1: Protein ORF6 protein / / Accessory protein 6 / Non-structural protein 6 / ns6 / Protein X3


Mass: 7533.843 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Severe acute respiratory syndrome coronavirus
Production host: Insect cell expression vector pTIE1 (others)
References: UniProt: P59634
#2: Protein mRNA export factor / Rae1 protein homolog / mRNA-associated protein mrnp 41


Mass: 41017.078 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: RAE1, MRNP41
Production host: Insect cell expression vector pTIE1 (others)
References: UniProt: P78406
#3: Protein Nuclear pore complex protein Nup98-Nup96 / Nuclear pore


Mass: 197780.547 Da / Num. of mol.: 2
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Homo sapiens (human) / Gene: NUP98, ADAR2
Production host: Insect cell expression vector pTIE1 (others)
References: UniProt: P52948, Hydrolases; Acting on peptide bonds (peptidases); Serine endopeptidases
#4: Water ChemComp-HOH / water / Water


Mass: 18.015 Da / Num. of mol.: 189 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION / Number of used crystals: 1

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Sample preparation

CrystalDensity Matthews: 2.32 Å3/Da / Density % sol: 0.47 %
Crystal growTemperature: 296 K / Method: vapor diffusion, hanging drop / pH: 5.5 / Details: 0.1m Bis-tris ph5.5,45%PEG4000

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Data collection

DiffractionMean temperature: 100 K / Serial crystal experiment: N
Diffraction sourceSource: SYNCHROTRON / Site: SSRF / Beamline: BL19U1 / Wavelength: 0.9785 Å
DetectorType: DECTRIS PILATUS3 S 6M / Detector: PIXEL / Date: Jun 27, 2021
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9785 Å / Relative weight: 1
ReflectionResolution: 2.39→47.874 Å / Num. obs: 60024 / % possible obs: 99.7 % / Redundancy: 3.49 % / Rmerge(I) obs: 0.131 / Net I/σ(I): 7.62
Reflection shellResolution: 2.39→2.54 Å / Rmerge(I) obs: 0.624 / Mean I/σ(I) obs: 2.01 / Num. unique obs: 9702 / % possible all: 99.2

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Processing

Software
NameVersionClassification
PHENIX1.18.2_3874refinement
PDB_EXTRACT3.27data extraction
XDSdata reduction
XDSdata scaling
Cootmodel building
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4OWR

4owr


Resolution: 2.39→47.87 Å / SU ML: 0.3 / Cross valid method: THROUGHOUT / σ(F): 1.34 / Phase error: 28.06 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2581 1599 5.22 %
Rwork0.2196 29033 -
obs0.2215 30632 99.68 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 166.7 Å2 / Biso mean: 44.0392 Å2 / Biso min: 20.47 Å2
Refinement stepCycle: final / Resolution: 2.39→47.87 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6099 0 0 189 6288
Biso mean---37.39 -
Num. residues----768
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 11

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
2.39-2.470.32091400.28382598273899
2.47-2.560.31781400.286326102750100
2.56-2.660.33261440.271126472791100
2.66-2.780.31031380.26326182756100
2.78-2.930.31731490.264726172766100
2.93-3.110.28821640.246626192783100
3.11-3.350.28851260.227726602786100
3.35-3.690.24551490.202826642813100
3.69-4.220.21011560.18826192775100
4.22-5.320.19231400.165726762816100
5.32-47.870.24991530.21827052858100
Refinement TLS params.Method: refined / Origin x: -23.9906 Å / Origin y: -2.745 Å / Origin z: 10.8391 Å
111213212223313233
T0.3507 Å20.0093 Å20.0194 Å2-0.2456 Å2-0.0612 Å2--0.221 Å2
L0.036 °20.0412 °20.0312 °2-1.7227 °2-0.1483 °2--0.1672 °2
S0.0575 Å °-0.0149 Å °0.0123 Å °-0.0324 Å °-0.0798 Å °0.2313 Å °-0.0473 Å °0.0193 Å °0.0055 Å °
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection detailsAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1allE53 - 62
2X-RAY DIFFRACTION1allF52 - 63
3X-RAY DIFFRACTION1allA31 - 365
4X-RAY DIFFRACTION1allC31 - 364
5X-RAY DIFFRACTION1allB158 - 213
6X-RAY DIFFRACTION1allD158 - 213
7X-RAY DIFFRACTION1allS1 - 189

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