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- PDB-5x50: RNA Polymerase II from Komagataella Pastoris (Type-2 crystal) -

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Basic information

Entry
Database: PDB / ID: 5x50
TitleRNA Polymerase II from Komagataella Pastoris (Type-2 crystal)
Components
  • (DNA-directed RNA polymerase ...Polymerase) x 3
  • (RNA polymerase II ...) x 4
  • (RNA polymerase subunit ...) x 4
  • RNA polymerase subunit, found in RNA polymerase complexes I, II, and III
KeywordsTRANSFERASE / transcription / RNA polymerase
Function / homology
Function and homology information


telomeric repeat-containing RNA transcription by RNA polymerase II / regulation of septum digestion after cytokinesis / heterochromatin assembly by small RNA / RNA polymerase III activity / recruitment of 3'-end processing factors to RNA polymerase II holoenzyme complex / mRNA export from nucleus in response to heat stress / termination of RNA polymerase II transcription / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / RNA polymerase II activity / RNA polymerase I activity ...telomeric repeat-containing RNA transcription by RNA polymerase II / regulation of septum digestion after cytokinesis / heterochromatin assembly by small RNA / RNA polymerase III activity / recruitment of 3'-end processing factors to RNA polymerase II holoenzyme complex / mRNA export from nucleus in response to heat stress / termination of RNA polymerase II transcription / nuclear-transcribed mRNA catabolic process, deadenylation-dependent decay / RNA polymerase II activity / RNA polymerase I activity / positive regulation of translational initiation / tRNA transcription by RNA polymerase III / transcription by RNA polymerase I / transcription by RNA polymerase III / mRNA cleavage / RNA polymerase I complex / RNA polymerase III complex / RNA polymerase II, core complex / pericentric heterochromatin / transcription initiation from RNA polymerase II promoter / translation initiation factor binding / P-body / ribonucleoside binding / DNA-directed 5'-3' RNA polymerase activity / transcription by RNA polymerase II / DNA-directed RNA polymerase / single-stranded DNA binding / single-stranded RNA binding / nucleic acid binding / RNA-directed 5'-3' RNA polymerase activity / transcription, DNA-templated / protein dimerization activity / chromatin / nucleotide binding / nucleolus / RNA binding / DNA binding / zinc ion binding / metal ion binding
Similarity search - Function
Rpb4/RPC9 superfamily / RNA polymerase Rpb4/RPC9, core / DNA-directed RNA-polymerase II subunit / RNA polymerase Rpb1, domain 7 / RNA polymerase Rpb1, domain 7 superfamily / RNA polymerase Rpb1, domain 7 / RNA polymerase Rpb1, domain 6 / RNA polymerase II, heptapeptide repeat, eukaryotic / RNA polymerase Rpb1 C-terminal repeat / Eukaryotic RNA polymerase II heptapeptide repeat. ...Rpb4/RPC9 superfamily / RNA polymerase Rpb4/RPC9, core / DNA-directed RNA-polymerase II subunit / RNA polymerase Rpb1, domain 7 / RNA polymerase Rpb1, domain 7 superfamily / RNA polymerase Rpb1, domain 7 / RNA polymerase Rpb1, domain 6 / RNA polymerase II, heptapeptide repeat, eukaryotic / RNA polymerase Rpb1 C-terminal repeat / Eukaryotic RNA polymerase II heptapeptide repeat. / RNA polymerase Rpb1, domain 6 / RNA polymerase RBP11 / Pol II subunit B9, C-terminal zinc ribbon / RNA polymerase subunit Rpb4/RPC9 / RNA polymerase Rpb4 / S1 domain profile. / RNA polymerase Rpb7-like , N-terminal / SHS2 domain found in N terminus of Rpb7p/Rpc25p/MJ0397 / HRDC-like superfamily / RNA polymerase Rpb2, domain 4 / RNA polymerase Rpb2, domain 4 / RNA polymerase Rpb7-like, N-terminal domain superfamily / RNA polymerase Rpb2, domain 5 / RNA polymerase Rpb2, domain 5 / RNA polymerases, subunit N, zinc binding site / RNA polymerases N / 8 kDa subunit / RNA polymerases N / 8 Kd subunits signature. / RNA polymerase subunit RPB10 / DNA-directed RNA polymerase subunit RPABC5/Rpb10 / RNA polymerase subunit 9 / DNA-directed RNA polymerase, M/15kDa subunit / RNA polymerases M/15 Kd subunit / DNA-directed RNA polymerases I, II, and III subunit RPABC4 / DNA-directed RNA polymerase M, 15kDa subunit, conserved site / RNA polymerases M / 15 Kd subunits signature. / RNA polymerase, Rpb5, N-terminal / DNA-directed RNA polymerase subunit Rpo5/Rpb5 / RNA polymerase Rpb5, N-terminal domain / RNA polymerase Rpb5, N-terminal domain superfamily / DNA-directed RNA polymerase subunit/transcription factor S / RNA polymerase, Rpb8 / RNA polymerase subunit 8 / RNA polymerase Rpb8 / DNA-directed RNA polymerase, subunit RPB6 / DNA directed RNA polymerase, 7 kDa subunit / RNA polymerase archaeal subunit P/eukaryotic subunit RPABC4 / RNA polymerase subunit CX / DNA-directed RNA polymerase, 30-40kDa subunit, conserved site / RNA polymerases D / 30 to 40 Kd subunits signature. / : / RNA polymerase Rpb5, C-terminal domain / RPB5-like RNA polymerase subunit superfamily / RNA polymerase Rpb3/Rpb11 dimerisation domain / : / RNA polymerases L / 13 to 16 Kd subunits signature. / RNA polymerase, subunit H/Rpb5, conserved site / RNA polymerase, subunit H/Rpb5 C-terminal / DNA-directed RNA polymerase, RBP11-like dimerisation domain / DNA-directed RNA polymerase Rpb11, 13-16kDa subunit, conserved site / RNA polymerases H / 23 Kd subunits signature. / RNA polymerase subunit RPABC4/transcription elongation factor Spt4 / DNA-directed RNA polymerase, 14-18kDa subunit, conserved site / Archaeal Rpo6/eukaryotic RPB6 RNA polymerase subunit / RNA polymerases K / 14 to 18 Kd subunits signature. / Zinc finger TFIIS-type profile. / Transcription factor S-II (TFIIS) / C2C2 Zinc finger / Zinc finger, TFIIS-type / S1 RNA binding domain / S1 domain / RNA polymerase Rpb1, domain 3 superfamily / RNA polymerase Rpb6 / RNA polymerase Rpb6 / RPB6/omega subunit-like superfamily / RNA polymerase, subunit omega/Rpo6/RPB6 / : / RNA polymerase Rpb2, domain 2 superfamily / RNA polymerase Rpb1, domain 2 / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 1 / RNA polymerase, N-terminal / RNA polymerase Rpb1, domain 4 / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, domain 3 / RNA polymerase Rpb1, funnel domain superfamily / RNA polymerase I subunit A N-terminus / RNA polymerase Rpb1, domain 1 / RNA polymerase Rpb1, domain 5 / RNA polymerase Rpb1, domain 4 / RNA polymerase, alpha subunit / RNA polymerase beta subunit / RNA polymerase Rpb2, domain 2 / RNA polymerase Rpb2, domain 2 / RNA polymerase, beta subunit, protrusion / RNA polymerases D / DNA-directed RNA polymerase, insert domain / DNA-directed RNA polymerase, insert domain superfamily / RNA polymerase Rpb3/Rpb11 dimerisation domain / RNA polymerase Rpb3/RpoA insert domain / DNA-directed RNA polymerase, RpoA/D/Rpb3-type
Similarity search - Domain/homology
DNA-directed RNA polymerases I, II, and III subunit RPABC1 / RNA polymerase II subunit / RNA polymerase II third largest subunit B44, part of central core / DNA-directed RNA polymerase subunit / RNA polymerase II subunit B12.5 / RNA polymerase subunit ABC23, common to RNA polymerases I, II, and III / RNA polymerase II subunit B32 / DNA-directed RNA polymerases I, II, and III subunit RPABC3 / RNA polymerase subunit ABC10-beta, common to RNA polymerases I, II, and III / DNA-directed RNA polymerase subunit beta ...DNA-directed RNA polymerases I, II, and III subunit RPABC1 / RNA polymerase II subunit / RNA polymerase II third largest subunit B44, part of central core / DNA-directed RNA polymerase subunit / RNA polymerase II subunit B12.5 / RNA polymerase subunit ABC23, common to RNA polymerases I, II, and III / RNA polymerase II subunit B32 / DNA-directed RNA polymerases I, II, and III subunit RPABC3 / RNA polymerase subunit ABC10-beta, common to RNA polymerases I, II, and III / DNA-directed RNA polymerase subunit beta / RNA polymerase subunit, found in RNA polymerase complexes I, II, and III / DNA-directed RNA polymerase subunit
Similarity search - Component
Biological speciesKomagataella phaffii (fungus)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / molecular replacement / Resolution: 4.293 Å
AuthorsEhara, H. / Umehara, T. / Sekine, S. / Yokoyama, S.
CitationJournal: Biochem. Biophys. Res. Commun. / Year: 2017
Title: Crystal structure of RNA polymerase II from Komagataella pastoris
Authors: Ehara, H. / Umehara, T. / Sekine, S.I. / Yokoyama, S.
History
DepositionFeb 14, 2017Deposition site: PDBJ / Processing site: PDBJ
Revision 1.0May 17, 2017Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: DNA-directed RNA polymerase subunit
B: DNA-directed RNA polymerase subunit beta
C: RNA polymerase II third largest subunit B44, part of central core
D: RNA polymerase II subunit B32
E: RNA polymerase subunit ABC27, common to RNA polymerases I, II, and III
F: RNA polymerase subunit ABC23, common to RNA polymerases I, II, and III
G: RNA polymerase II subunit
H: RNA polymerase subunit ABC14.5, common to RNA polymerases I, II, and III
I: DNA-directed RNA polymerase subunit
J: RNA polymerase subunit ABC10-beta, common to RNA polymerases I, II, and III
K: RNA polymerase II subunit B12.5
L: RNA polymerase subunit, found in RNA polymerase complexes I, II, and III
hetero molecules


Theoretical massNumber of molelcules
Total (without water)511,00620
Polymers510,48212
Non-polymers5238
Water0
1


  • Idetical with deposited unit
  • defined by author&software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area56970 Å2
ΔGint-339 kcal/mol
Surface area167070 Å2
MethodPISA
Unit cell
γ
α
β
Length a, b, c (Å)211.750, 211.750, 132.280
Angle α, β, γ (deg.)90.000, 90.000, 120.000
Int Tables number144
Space group name H-MP31

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Components

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DNA-directed RNA polymerase ... , 3 types, 3 molecules ABI

#1: Protein DNA-directed RNA polymerase subunit / Polymerase


Mass: 194107.422 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R4Y0, DNA-directed RNA polymerase
#2: Protein DNA-directed RNA polymerase subunit beta / Polymerase


Mass: 139746.094 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4QZQ7, DNA-directed RNA polymerase
#9: Protein DNA-directed RNA polymerase subunit / Polymerase


Mass: 13612.320 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain ATCC 76273 / CBS 7435 / CECT 11047 / NRRL Y-11430 / Wegner 21-1) (fungus)
Strain: ATCC 76273 / CBS 7435 / CECT 11047 / NRRL Y-11430 / Wegner 21-1
References: UniProt: F2QPE6

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RNA polymerase II ... , 4 types, 4 molecules CDGK

#3: Protein RNA polymerase II third largest subunit B44, part of central core


Mass: 34216.293 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R7L2
#4: Protein RNA polymerase II subunit B32 /


Mass: 20622.980 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R2U9
#7: Protein RNA polymerase II subunit /


Mass: 18802.625 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R9A1
#11: Protein RNA polymerase II subunit B12.5 /


Mass: 13832.896 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R3Z5

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RNA polymerase subunit ... , 4 types, 4 molecules EFHJ

#5: Protein RNA polymerase subunit ABC27, common to RNA polymerases I, II, and III /


Mass: 24962.680 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R3P8
#6: Protein RNA polymerase subunit ABC23, common to RNA polymerases I, II, and III /


Mass: 17803.588 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R1V1
#8: Protein RNA polymerase subunit ABC14.5, common to RNA polymerases I, II, and III /


Mass: 16249.220 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R273
#10: Protein RNA polymerase subunit ABC10-beta, common to RNA polymerases I, II, and III /


Mass: 8554.064 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4R009

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Protein / Non-polymers , 2 types, 9 molecules L

#12: Protein RNA polymerase subunit, found in RNA polymerase complexes I, II, and III /


Mass: 7972.201 Da / Num. of mol.: 1 / Source method: isolated from a natural source
Source: (natural) Komagataella phaffii (strain GS115 / ATCC 20864) (fungus)
Strain: GS115 / ATCC 20864 / References: UniProt: C4QWA8
#13: Chemical
ChemComp-ZN / ZINC ION


Mass: 65.409 Da / Num. of mol.: 8 / Source method: obtained synthetically / Formula: Zn

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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: 3.35 Å3/Da / Density % sol: 63.33 %
Crystal growTemperature: 287 K / Method: vapor diffusion, sitting drop
Details: 0.2 M sodium citrate, 8% PGA-LM and 1 % dextran sulfate sodium salt

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SPring-8 / Beamline: BL41XU / Wavelength: 1 Å
DetectorType: RAYONIX MX225HE / Detector: CCD / Date: Oct 28, 2011
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 1 Å / Relative weight: 1
ReflectionResolution: 4.29→40.704 Å / Num. obs: 45119 / % possible obs: 99.6 % / Observed criterion σ(I): -3 / Redundancy: 4.857 % / Biso Wilson estimate: 191.58 Å2 / CC1/2: 0.994 / Rmerge(I) obs: 0.188 / Rrim(I) all: 0.211 / Χ2: 0.976 / Net I/σ(I): 5.97 / Num. measured all: 219141
Reflection shell

Diffraction-ID: 1

Resolution (Å)Redundancy (%)Rmerge(I) obsMean I/σ(I) obsNum. measured obsNum. possibleNum. unique obsCC1/2Rrim(I) all% possible all
4.29-4.554.7661.4790.9833980728671300.4571.66297.9
4.55-4.864.9250.9981.633662683568350.6681.118100
4.86-5.254.9290.762.1331437637963780.7530.851100
5.25-5.744.9370.6122.6929129590059000.8060.685100
5.74-6.414.9440.4313.8826333532653260.8930.482100
6.41-7.384.9190.2237.0323115469946990.970.25100
7.38-8.994.8260.09514.8119055394839480.9930.106100
8.99-12.54.6770.06121.914612312431240.9950.069100
12.5-40.7044.3950.05723.317818182317790.9940.06497.6

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Phasing

PhasingMethod: molecular replacement

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Processing

Software
NameVersionClassification
PHENIX1.10.1_2155refinement
XSCALEdata scaling
PDB_EXTRACT3.22data extraction
XDSdata reduction
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 2VUM

2vum


Resolution: 4.293→40.704 Å / SU ML: 0.78 / Cross valid method: FREE R-VALUE / σ(F): 1.77 / Phase error: 33.15 / Stereochemistry target values: ML
RfactorNum. reflection% reflection
Rfree0.2957 1988 4.41 %
Rwork0.2353 43093 -
obs0.238 45081 99.57 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Displacement parametersBiso max: 477.15 Å2 / Biso mean: 223.7058 Å2 / Biso min: 46.79 Å2
Refinement stepCycle: final / Resolution: 4.293→40.704 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms28430 0 8 0 28438
Biso mean--257.83 --
Num. residues----3829
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.00628881
X-RAY DIFFRACTIONf_angle_d1.69638973
X-RAY DIFFRACTIONf_chiral_restr0.5134478
X-RAY DIFFRACTIONf_plane_restr0.0085013
X-RAY DIFFRACTIONf_dihedral_angle_d18.90710563
LS refinement shell

Refine-ID: X-RAY DIFFRACTION / Rfactor Rfree error: 0 / Total num. of bins used: 14

Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkNum. reflection all% reflection obs (%)
4.2934-4.40070.38461370.33442974311195
4.4007-4.51950.34851460.280530583204100
4.5195-4.65230.2931380.265630623200100
4.6523-4.80230.37881460.255931363282100
4.8023-4.97360.30551380.26930743212100
4.9736-5.17240.33511420.256630863228100
5.1724-5.40730.31241440.255430793223100
5.4073-5.69170.35091400.24131213261100
5.6917-6.04720.30271380.234730853223100
6.0472-6.51240.34261480.241830943242100
6.5124-7.16450.33481400.232330693209100
7.1645-8.19390.3061380.212431083246100
8.1939-10.29570.21191480.188430833231100
10.2957-40.70620.27661450.23613064320999

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