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Title:Three-dimensional structure of the RNA polymerase II-Iwr1 complex
Authors:Czeko E, Seizl M, Augsberger C, Mielke T, Cramer P
Sample:RNA polymerase II-Iwr1
Method:Single particle reconstruction (20.9 angstroms resolution)
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Sample
Sample name: RNA polymerase II-Iwr1
Oligomeric state: Monomeric
Theoretical molecular weight of the sample: 0.55
Components:
ID Type Name Exp. MW (MDa) Oligomeric details Recombinant expression Synthetic Organism UniProt identifier GO identifier InterPro identifier Virus identifier Details
1proteinDNA-directed RNA polymerasefalseSaccharomyces cerevisiae
2proteinImport adaptorMonomertrueSaccharomyces cerevisiae
Experiment
Specimen state: Particle
Specimen preparation:
pHSpecimen conc.DetailsStainingSpecimen support details
7.250.1 mg/mL5 mM HEPES, 40 mM Ammonium sulfate, 10 microM Zinc chloride, 10 mM DTT
Vitrification:
Cryogen nameHumidityTemp.Instr.MethodTime resolvedDetails
ETHANE95%80 KFEI VITROBOTBlot for 10 s before plunging msVitrification instrument: FEI Vitrobot
Imaging:
MicroscopeVoltageIllumination modeImaging modeCsDefocus min.Defocus max.Nominal mag.Calibrated mag.Electron sourceDetectorDetector distanceAstigmatism
FEI POLARA 300300 kVSPOT SCANBRIGHT FIELD2.0 mm1500 nm3500 nm67000FIELD EMISSION GUNKODAK SO-163 FILM mm

Specimen holderHolder modelTilt min.Tilt max.Energy filterEnergy windowTemp.Temp. min.Temp. max.Beam tiltElectron doseOther detailsDate
Multispecimen holderOTHER°° eV80 K K K mrad25 e/Å2
Processing
Protocol:Reference projection matching
Software:SPIDER
CTF correction:Defocus groups
Number of particles:25206
Imposed symmetry:C1
Resolution by author:20.9 Å
Resolution method:FSC 0.5
Processing details:Particles were selected using Signature and thereafter checked manually.
Scanned images:
Num. imagesSampling sizeOD rangeQuant. bit numberOther detailsScanner
35 μm/pixelOTHER
Fitting:
PDBProtocolTarget crit.SoftwareB valueFitting spacePDB chainDetails
1WCM rigid bodyLeast-squareChimeraREALProtocol: Rigid body