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- PDB-5t94: Crystal structure of Kap60 bound to yeast RCC1 (Prp20) -

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Basic information

Entry
Database: PDB / ID: 5t94
TitleCrystal structure of Kap60 bound to yeast RCC1 (Prp20)
Components
  • Guanine nucleotide exchange factor SRM1
  • Importin subunit alpha
KeywordsPROTEIN TRANSPORT / NUCLEAR PROTEIN / Nuclear Import / Importin alpha / RCC1 like Domain / RLD / NLS / PROTEIN TRANSPORT - NUCLEAR PROTEIN complex
Function / homology
Function and homology information


proteasome localization / response to pheromone / Postmitotic nuclear pore complex (NPC) reformation / import into nucleus / NLS-dependent protein nuclear import complex / protein targeting to membrane / nuclear import signal receptor activity / poly(A)+ mRNA export from nucleus / nuclear localization sequence binding / NLS-bearing protein import into nucleus ...proteasome localization / response to pheromone / Postmitotic nuclear pore complex (NPC) reformation / import into nucleus / NLS-dependent protein nuclear import complex / protein targeting to membrane / nuclear import signal receptor activity / poly(A)+ mRNA export from nucleus / nuclear localization sequence binding / NLS-bearing protein import into nucleus / nucleus organization / ribosomal subunit export from nucleus / guanyl-nucleotide exchange factor activity / protein import into nucleus / disordered domain specific binding / nuclear envelope / cell cycle / cell division / protein-containing complex binding / chromatin / perinuclear region of cytoplasm / protein-containing complex / nucleoplasm / nucleus / cytoplasm / cytosol
Similarity search - Function
Regulator of chromosome condensation (RCC1) signature 1. / Regulator of chromosome condensation (RCC1) repeat / Regulator of chromosome condensation 1/beta-lactamase-inhibitor protein II / Regulator of chromosome condensation (RCC1) signature 2. / Regulator of chromosome condensation (RCC1) repeat / Regulator of chromosome condensation, RCC1 / Regulator of chromosome condensation (RCC1) repeat profile. / Regulator of chromosome condensation 1/beta-lactamase-inhibitor protein II / Importin subunit alpha / Atypical Arm repeat ...Regulator of chromosome condensation (RCC1) signature 1. / Regulator of chromosome condensation (RCC1) repeat / Regulator of chromosome condensation 1/beta-lactamase-inhibitor protein II / Regulator of chromosome condensation (RCC1) signature 2. / Regulator of chromosome condensation (RCC1) repeat / Regulator of chromosome condensation, RCC1 / Regulator of chromosome condensation (RCC1) repeat profile. / Regulator of chromosome condensation 1/beta-lactamase-inhibitor protein II / Importin subunit alpha / Atypical Arm repeat / Importin-alpha, importin-beta-binding domain superfamily / Importin beta binding domain / Atypical Arm repeat / Importin-alpha, importin-beta-binding domain / IBB domain profile. / Armadillo/plakoglobin ARM repeat profile. / Armadillo/beta-catenin-like repeat / Armadillo/beta-catenin-like repeats / Armadillo / Leucine-rich Repeat Variant / Leucine-rich Repeat Variant / 7 Propeller / Methylamine Dehydrogenase; Chain H / Armadillo-like helical / Alpha Horseshoe / Armadillo-type fold / Mainly Beta / Mainly Alpha
Similarity search - Domain/homology
Guanine nucleotide exchange factor SRM1 / Importin subunit alpha
Similarity search - Component
Biological speciesSaccharomyces cerevisiae (brewer's yeast)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 2.631 Å
AuthorsSankhala, R.S. / Lokareddy, R.K. / Pumroy, R.A. / Cingolani, G.
Funding support United States, 2items
OrganizationGrant numberCountry
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM074846 United States
National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)GM100888 United States
CitationJournal: Nat Commun / Year: 2017
Title: Three-dimensional context rather than NLS amino acid sequence determines importin alpha subtype specificity for RCC1.
Authors: Sankhala, R.S. / Lokareddy, R.K. / Begum, S. / Pumroy, R.A. / Gillilan, R.E. / Cingolani, G.
History
DepositionSep 9, 2016Deposition site: RCSB / Processing site: RCSB
Revision 1.0Sep 13, 2017Provider: repository / Type: Initial release
Revision 1.1Nov 1, 2017Group: Author supporting evidence / Database references
Category: citation / citation_author / pdbx_struct_assembly_auth_evidence
Item: _citation.country / _citation.journal_abbrev ..._citation.country / _citation.journal_abbrev / _citation.journal_id_CSD / _citation.journal_id_ISSN / _citation.journal_volume / _citation.page_first / _citation.page_last / _citation.pdbx_database_id_DOI / _citation.pdbx_database_id_PubMed / _citation.title / _citation.year
Revision 1.2Apr 17, 2019Group: Author supporting evidence / Data collection / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.3Dec 25, 2019Group: Author supporting evidence / Category: pdbx_audit_support / Item: _pdbx_audit_support.funding_organization
Revision 1.4Oct 4, 2023Group: Data collection / Database references / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / pdbx_initial_refinement_model
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Guanine nucleotide exchange factor SRM1
B: Importin subunit alpha


Theoretical massNumber of molelcules
Total (without water)113,5822
Polymers113,5822
Non-polymers00
Water2,036113
1


  • Idetical with deposited unit
  • defined by author&software
  • Evidence: gel filtration, SAXS
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3780 Å2
ΔGint-2 kcal/mol
Surface area36180 Å2
MethodPISA
Unit cell
Length a, b, c (Å)91.050, 99.200, 125.030
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121
DetailsDimer as determined by Gel filtration chromatography and further verified using small angle x-ray scattering method.

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Components

#1: Protein Guanine nucleotide exchange factor SRM1 / Pheromone response pathway component SRM1 / Pre-mRNA-processing protein 20 / Regulator of ...Pheromone response pathway component SRM1 / Pre-mRNA-processing protein 20 / Regulator of chromosome condensation / Suppressor of receptor mutations 1 / mRNA transport protein 1


Mass: 53082.684 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Details: Ran guanyl-nucleotide exchange factor
Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast)
Strain: ATCC 204508 / S288c / Gene: SRM1, MTR1, PRP20, YGL097W / Plasmid: pGEX-6P-1 / Production host: Escherichia coli (E. coli) / References: UniProt: P21827
#2: Protein Importin subunit alpha / Karyopherin subunit alpha / Karyopherin-60 / Serine-rich RNA polymerase I suppressor protein


Mass: 60499.043 Da / Num. of mol.: 1 / Mutation: T205A, D276E, K400R
Source method: isolated from a genetically manipulated source
Details: Yeast nuclear import adapter protein
Source: (gene. exp.) Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (yeast)
Strain: ATCC 204508 / S288c / Gene: SRP1, KAP60, YNL189W, N1606 / Plasmid: pET28a / Production host: Escherichia coli (E. coli) / References: UniProt: Q02821
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 113 / 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: 2.49 Å3/Da / Density % sol: 50.52 % / Description: Large thick block
Crystal growTemperature: 289 K / Method: vapor diffusion, hanging drop
Details: 0.1 M Na cacodylate, 0.1 M NaCl and 1.1 M ammonium sulfate

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: SSRL / Beamline: BL14-1 / Wavelength: 0.97 Å
DetectorType: RAYONIX MX325HE / Detector: CCD / Date: Jul 16, 2016
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.97 Å / Relative weight: 1
ReflectionResolution: 2.63→15 Å / Num. obs: 33934 / % possible obs: 96 % / Redundancy: 4.7 % / Biso Wilson estimate: 72.8 Å2 / Rsym value: 0.049 / Net I/σ(I): 49.9
Reflection shellResolution: 2.63→2.72 Å / Redundancy: 2.7 % / Rmerge(I) obs: 0.644 / Mean I/σ(I) obs: 3.4 / % possible all: 85.8

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Processing

Software
NameVersionClassification
PHENIX1.10.1-2155refinement
HKL-2000data reduction
HKL-2000data scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT
Starting model: 4PVZ and 3OF7
Resolution: 2.631→14.894 Å / SU ML: 0.46 / Cross valid method: FREE R-VALUE / σ(F): 1.35 / Phase error: 32.61
RfactorNum. reflection% reflection
Rfree0.2589 1540 4.89 %
Rwork0.2167 --
obs0.2188 31489 92.42 %
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å
Displacement parametersBiso mean: 84.5 Å2
Refinement stepCycle: LAST / Resolution: 2.631→14.894 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms6808 0 0 113 6921
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0026941
X-RAY DIFFRACTIONf_angle_d0.4969414
X-RAY DIFFRACTIONf_dihedral_angle_d14.9934210
X-RAY DIFFRACTIONf_chiral_restr0.0391063
X-RAY DIFFRACTIONf_plane_restr0.0041226
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.6311-2.71550.3944690.41181404X-RAY DIFFRACTION48
2.7155-2.8120.4641030.38772306X-RAY DIFFRACTION79
2.812-2.92370.39751370.36842822X-RAY DIFFRACTION96
2.9237-3.05570.40731230.32952842X-RAY DIFFRACTION97
3.0557-3.21530.36681320.29732910X-RAY DIFFRACTION99
3.2153-3.41450.30821650.25612882X-RAY DIFFRACTION99
3.4145-3.67440.26131520.21992923X-RAY DIFFRACTION99
3.6744-4.03750.23211540.18352894X-RAY DIFFRACTION99
4.0375-4.60650.18761630.15752945X-RAY DIFFRACTION100
4.6065-5.74770.21321820.17512956X-RAY DIFFRACTION99
5.7477-14.89390.22721600.17243065X-RAY DIFFRACTION99
Refinement TLS params.

Method: refined / Refine-ID: X-RAY DIFFRACTION

IDL112)L122)L132)L222)L232)L332)S11 (Å °)S12 (Å °)S13 (Å °)S21 (Å °)S22 (Å °)S23 (Å °)S31 (Å °)S32 (Å °)S33 (Å °)T112)T122)T132)T222)T232)T332)Origin x (Å)Origin y (Å)Origin z (Å)
10.31390.50320.41911.03810.21860.5993-0.0554-0.7775-0.924-0.2215-0.0478-0.11970.3572-0.54160.0040.34170.04620.01870.5827-0.0120.476219.4398103.7229-0.9316
23.94640.2034-0.03372.7781-0.13582.4940.0829-0.6807-0.43520.3425-0.15060.10070.4757-0.1666-0.00460.29210.010.01650.55510.13040.229725.0257111.747114.666
35.13290.206-0.22454.01810.16192.9226-0.02310.59340.4264-0.2547-0.0517-0.2666-0.24480.2784-0.00570.1210.05980.01430.45660.06090.286437.7198124.17390.7874
42.87260.60411.672.79580.52652.33750.42250.6473-1.0869-0.14310.1098-0.15250.73830.1661-0.19390.5010.019-0.10070.4377-0.28440.61329.611995.806-13.9251
50.2184-0.1342-0.37391.5437-0.09240.72971.1589-1.1669-1.07570.0228-0.15860.09190.6256-0.94530.00831.068-0.772-0.55811.36050.42871.1945-32.397578.6453-14.2015
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'A' and (resid 2 through 64 )
2X-RAY DIFFRACTION2chain 'A' and (resid 65 through 331 )
3X-RAY DIFFRACTION3chain 'A' and (resid 332 through 482 )
4X-RAY DIFFRACTION4chain 'B' and (resid 88 through 284 )
5X-RAY DIFFRACTION5chain 'B' and (resid 285 through 509 )

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