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- PDB-2euz: Structure of a Ndt80-DNA complex (MSE mutant mC5T) -

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Basic information

Entry
Database: PDB / ID: 2euz
TitleStructure of a Ndt80-DNA complex (MSE mutant mC5T)
Components
  • 5'-D(*AP*GP*TP*TP*TP*TP*TP*AP*TP*GP*TP*CP*GP*C)-3'
  • 5'-D(*TP*GP*CP*GP*AP*CP*AP*TP*AP*AP*AP*AP*AP*C)-3'
  • NDT80 protein
KeywordsCELL CYCLE/DNA / BETA-BARREL / IG-FOLD TRANSCRIPTION FACTOR / CELL CYCLE-DNA COMPLEX
Function / homology
Function and homology information


sporulation / nuclear chromosome / meiotic cell cycle / sequence-specific DNA binding / DNA-binding transcription factor activity / cell division / positive regulation of transcription by RNA polymerase II
Similarity search - Function
NDT80 DNA-binding domain / : / NDT80 DNA-binding domain / NDT80 DNA-binding domain superfamily / NDT80 / PhoG like DNA-binding family / NDT80 DNA-binding domain profile. / p53-like transcription factor, DNA-binding / Immunoglobulin-like / Sandwich / Mainly Beta
Similarity search - Domain/homology
DNA / DNA (> 10) / Meiosis-specific transcription factor NDT80
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
MethodX-RAY DIFFRACTION / SYNCHROTRON / FOURIER SYNTHESIS / Resolution: 1.56 Å
AuthorsLamoureux, J.S. / Glover, J.N.
CitationJournal: Structure / Year: 2006
Title: Principles of Protein-DNA Recognition Revealed in the Structural Analysis of Ndt80-MSE DNA Complexes.
Authors: Lamoureux, J.S. / Glover, J.N.
History
DepositionOct 30, 2005Deposition site: RCSB / Processing site: RCSB
Revision 1.0Mar 21, 2006Provider: repository / Type: Initial release
Revision 1.1May 1, 2008Group: Version format compliance
Revision 1.2Jul 13, 2011Group: Version format compliance
Revision 1.3Oct 20, 2021Group: Database references / Category: database_2 / struct_ref_seq_dif
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ref_seq_dif.details
Revision 1.4Aug 23, 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
B: 5'-D(*TP*GP*CP*GP*AP*CP*AP*TP*AP*AP*AP*AP*AP*C)-3'
C: 5'-D(*AP*GP*TP*TP*TP*TP*TP*AP*TP*GP*TP*CP*GP*C)-3'
A: NDT80 protein


Theoretical massNumber of molelcules
Total (without water)48,1493
Polymers48,1493
Non-polymers00
Water6,702372
1


  • Idetical with deposited unit
  • defined by author
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Unit cell
Length a, b, c (Å)70.282, 78.839, 161.651
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number20
Space group name H-MC2221

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Components

#1: DNA chain 5'-D(*TP*GP*CP*GP*AP*CP*AP*TP*AP*AP*AP*AP*AP*C)-3'


Mass: 4281.832 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: MUTANT MSE DNA STRAND 1
#2: DNA chain 5'-D(*AP*GP*TP*TP*TP*TP*TP*AP*TP*GP*TP*CP*GP*C)-3'


Mass: 4276.786 Da / Num. of mol.: 1 / Source method: obtained synthetically / Details: MUTANT MSE DNA STRAND 2
#3: Protein NDT80 protein


Mass: 39590.527 Da / Num. of mol.: 1 / Fragment: NDT80 DNA-binding domain / Mutation: s146g, i200t
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Saccharomyces cerevisiae (brewer's yeast)
Gene: NDT80 / Plasmid: pGEX-6P1 / Species (production host): Escherichia coli / Production host: Escherichia coli BL21 (bacteria) / Strain (production host): BL21 / References: UniProt: P38830
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 372 / 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: 2.31 Å3/Da / Density % sol: 46.75 %
Crystal growTemperature: 295 K / Method: vapor diffusion, hanging drop / pH: 7
Details: 30% PEG 400, 50 mM bis-tris-propane pH 7.0, 100 mM NaCl, 50 mM CaCl2, and 2 mM DTT. 1:1 Molar ratio protein:DNA, protein at 10mg/ml , VAPOR DIFFUSION, HANGING DROP, temperature 295K
Components of the solutions
IDNameCrystal-IDSol-ID
1PEG 40011
2bis-tris-propane11
3NaCl11
4CaCl211
5DTT11
6H2O11
7PEG 40012
8NaCl12
9CaCl212

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Data collection

DiffractionMean temperature: 105 K
Diffraction sourceSource: SYNCHROTRON / Site: ALS / Beamline: 8.3.1 / Wavelength: 1.072158 Å
DetectorType: ADSC QUANTUM 210 / Detector: CCD / Date: Jul 14, 2003 / Details: MIRROR
RadiationMonochromator: KOHZU: Double Crystal Si(111) / Protocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1.072158 Å / Relative weight: 1
ReflectionResolution: 1.56→23.55 Å / Num. all: 61653 / Num. obs: 61653 / % possible obs: 96.4 % / Observed criterion σ(F): 0 / Observed criterion σ(I): 0 / Redundancy: 4.6 % / Rmerge(I) obs: 0.051 / Rsym value: 0.051 / Net I/σ(I): 8.5
Reflection shell
Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. unique allRsym value% possible all
1.56-1.643.10.3412.180060.34186.9
1.64-1.743.50.2133.483040.21395.4
1.74-1.863.60.1285.480190.12897.4
1.86-2.013.70.1025.275520.10297.9
2.01-2.214.30.1035.469840.10398.3
2.21-2.475.30.0926.363260.09298.7
2.47-2.856.80.0778.156430.07799.2
2.85-3.4970.0511.748500.0599.7
3.49-4.936.70.03815.838150.03899.9
4.93-23.5560.03317.521540.03398.8

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Processing

Software
NameVersionClassificationNB
SCALAdata scaling
REFMACrefinement
PDB_EXTRACT1.701data extraction
MOSFLMdata reduction
CCP4(SCALA)data scaling
RefinementMethod to determine structure: FOURIER SYNTHESIS
Starting model: PDB 1MNN - MINUS DNA AT SEQUENCE CHANGES AND 1 BASEPAIR ADJACENT TO THOSE CHANGES

1mnn


Resolution: 1.56→23.55 Å / Cor.coef. Fo:Fc: 0.969 / Cor.coef. Fo:Fc free: 0.962 / SU B: 1.358 / SU ML: 0.048 / Cross valid method: THROUGHOUT / σ(F): 0 / σ(I): 0 / ESU R: 0.077 / ESU R Free: 0.074 / Stereochemistry target values: MAXIMUM LIKELIHOOD / Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS
RfactorNum. reflection% reflectionSelection details
Rfree0.194 3092 5 %RANDOM
Rwork0.176 ---
all0.176 61559 --
obs0.176 61559 96.04 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: BABINET MODEL WITH MASK
Displacement parametersBiso mean: 19.647 Å2
Baniso -1Baniso -2Baniso -3
1-0 Å20 Å20 Å2
2--0 Å20 Å2
3---0 Å2
Refinement stepCycle: LAST / Resolution: 1.56→23.55 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms2395 551 0 372 3318
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.0213068
X-RAY DIFFRACTIONr_bond_other_d0.0020.022488
X-RAY DIFFRACTIONr_angle_refined_deg1.4212.1894273
X-RAY DIFFRACTIONr_angle_other_deg0.91135868
X-RAY DIFFRACTIONr_dihedral_angle_1_deg6.6945290
X-RAY DIFFRACTIONr_chiral_restr0.0810.2456
X-RAY DIFFRACTIONr_gen_planes_refined0.0060.022942
X-RAY DIFFRACTIONr_gen_planes_other0.0020.02503
X-RAY DIFFRACTIONr_nbd_refined0.1960.3466
X-RAY DIFFRACTIONr_nbd_other0.2370.32941
X-RAY DIFFRACTIONr_nbtor_other0.0860.51679
X-RAY DIFFRACTIONr_xyhbond_nbd_refined0.2030.5470
X-RAY DIFFRACTIONr_symmetry_vdw_refined0.2070.39
X-RAY DIFFRACTIONr_symmetry_vdw_other0.2650.336
X-RAY DIFFRACTIONr_symmetry_hbond_refined0.4210.524
X-RAY DIFFRACTIONr_mcbond_it1.47631471
X-RAY DIFFRACTIONr_mcangle_it2.55942419
X-RAY DIFFRACTIONr_scbond_it3.02261597
X-RAY DIFFRACTIONr_scangle_it4.458101853
LS refinement shellResolution: 1.56→1.6 Å / Total num. of bins used: 20
RfactorNum. reflection
Rfree0.3 198
Rwork0.266 3696
obs-3894

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