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- PDB-4am6: C-TERMINAL DOMAIN OF ACTIN-RELATED PROTEIN ARP8 FROM S. CEREVISIAE -

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Basic information

Entry
Database: PDB / ID: 4am6
TitleC-TERMINAL DOMAIN OF ACTIN-RELATED PROTEIN ARP8 FROM S. CEREVISIAE
ComponentsACTIN-LIKE PROTEIN ARP8
KeywordsNUCLEAR PROTEIN / CHROMATIN REMODELLING COMPLEX / ATP-BINDING PROTEIN / NUCLEAR ACTIN-RELATED PROTEIN / TRANSCRIPTION REGULATION / DNA REPAIR
Function / homology
Function and homology information


Ino80 complex / mitotic recombination / double-strand break repair / cytoskeleton / chromatin remodeling / DNA repair / mRNA binding / DNA damage response / regulation of DNA-templated transcription / ATP binding ...Ino80 complex / mitotic recombination / double-strand break repair / cytoskeleton / chromatin remodeling / DNA repair / mRNA binding / DNA damage response / regulation of DNA-templated transcription / ATP binding / nucleus / cytoplasm
Similarity search - Function
Nucleotidyltransferase; domain 5 - #580 / ATPase, substrate binding domain, subdomain 4 / Actin; Chain A, domain 4 / ATPase, nucleotide binding domain / Actin / Actin family / Actin / ATPase, nucleotide binding domain / Nucleotidyltransferase; domain 5 / Alpha-Beta Complex ...Nucleotidyltransferase; domain 5 - #580 / ATPase, substrate binding domain, subdomain 4 / Actin; Chain A, domain 4 / ATPase, nucleotide binding domain / Actin / Actin family / Actin / ATPase, nucleotide binding domain / Nucleotidyltransferase; domain 5 / Alpha-Beta Complex / 2-Layer Sandwich / Alpha Beta
Similarity search - Domain/homology
Actin-like protein ARP8
Similarity search - Component
Biological speciesSACCHAROMYCES CEREVISIAE (brewer's yeast)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 2.7 Å
AuthorsWuerges, J. / Saravanan, M. / Bose, D. / Cook, N.J. / Zhang, X. / Wigley, D.B.
CitationJournal: Proc Natl Acad Sci U S A / Year: 2012
Title: Interactions between the nucleosome histone core and Arp8 in the INO80 chromatin remodeling complex.
Authors: Matheshwaran Saravanan / Jochen Wuerges / Daniel Bose / Elizabeth A McCormack / Nicola J Cook / Xiaodong Zhang / Dale B Wigley /
Abstract: Actin-related protein Arp8 is a component of the INO80 chromatin remodeling complex. Yeast Arp8 (yArp8) comprises two domains: a 25-KDa N-terminal domain, found only in yeast, and a 75-KDa C-terminal ...Actin-related protein Arp8 is a component of the INO80 chromatin remodeling complex. Yeast Arp8 (yArp8) comprises two domains: a 25-KDa N-terminal domain, found only in yeast, and a 75-KDa C-terminal domain (yArp8CTD) that contains the actin fold and is conserved across other species. The crystal structure shows that yArp8CTD contains three insertions within the actin core. Using a combination of biochemistry and EM, we show that Arp8 forms a complex with nucleosomes, and that the principal interactions are via the H3 and H4 histones, mediated through one of the yArp8 insertions. We show that recombinant yArp8 exists in monomeric and dimeric states, but the dimer is the biologically relevant form required for stable interactions with histones that exploits the twofold symmetry of the nucleosome core. Taken together, these data provide unique insight into the stoichiometry, architecture, and molecular interactions between components of the INO80 remodeling complex and nucleosomes, providing a first step toward building up the structure of the complex.
History
DepositionMar 7, 2012Deposition site: PDBE / Processing site: PDBE
Revision 1.0Dec 12, 2012Provider: repository / Type: Initial release
Revision 1.1Jan 16, 2013Group: Database references / Structure summary
Revision 1.2Mar 6, 2013Group: Data collection
Revision 1.3Nov 13, 2019Group: Data collection / Database references / Other / Category: pdbx_database_related / pdbx_database_status
Item: _pdbx_database_related.content_type / _pdbx_database_status.status_code_sf
Revision 1.4May 8, 2024Group: Data collection / Database references ...Data collection / Database references / Derived calculations / Refinement description
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / struct_ncs_dom_lim / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_ncs_dom_lim.beg_auth_comp_id / _struct_ncs_dom_lim.beg_label_asym_id / _struct_ncs_dom_lim.beg_label_comp_id / _struct_ncs_dom_lim.beg_label_seq_id / _struct_ncs_dom_lim.end_auth_comp_id / _struct_ncs_dom_lim.end_label_asym_id / _struct_ncs_dom_lim.end_label_comp_id / _struct_ncs_dom_lim.end_label_seq_id / _struct_site.pdbx_auth_asym_id / _struct_site.pdbx_auth_comp_id / _struct_site.pdbx_auth_seq_id

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: ACTIN-LIKE PROTEIN ARP8
B: ACTIN-LIKE PROTEIN ARP8
hetero molecules


Theoretical massNumber of molelcules
Total (without water)149,8194
Polymers149,6272
Non-polymers1922
Water57632
1
A: ACTIN-LIKE PROTEIN ARP8
hetero molecules


Theoretical massNumber of molelcules
Total (without water)75,0063
Polymers74,8131
Non-polymers1922
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
2
B: ACTIN-LIKE PROTEIN ARP8


Theoretical massNumber of molelcules
Total (without water)74,8131
Polymers74,8131
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
MethodPISA
Unit cell
Length a, b, c (Å)138.200, 87.859, 149.370
Angle α, β, γ (deg.)90.00, 115.40, 90.00
Int Tables number5
Space group name H-MC121
Noncrystallographic symmetry (NCS)NCS domain:
IDEns-IDDetails
11A
21B

NCS domain segments:

Ens-ID: 1

Dom-IDComponent-IDBeg auth comp-IDBeg label comp-IDEnd auth comp-IDEnd label comp-IDRefine codeAuth asym-IDLabel asym-IDAuth seq-IDLabel seq-ID
11ASNASNTRPTRP5AA263 - 30137 - 75
21ASNASNTRPTRP5BB263 - 30137 - 75
12LEULEUGLUGLU6AA302 - 31876 - 92
22LEULEUGLUGLU6BB302 - 31876 - 92
13GLNGLNMETMET5AA319 - 33893 - 112
23GLNGLNMETMET5BB319 - 33893 - 112
14ARGARGARGARG6AA339 - 391113 - 165
24ARGARGARGARG6BB339 - 391113 - 165
15CYSCYSPROPRO5AA392 - 442166 - 216
25CYSCYSPROPRO5BB392 - 442166 - 216
16THRTHRCYSCYS4AA443 - 489217 - 263
26THRTHRCYSCYS4BB443 - 489217 - 263
17CYSCYSSERSER4AA499 - 555273 - 329
27CYSCYSSERSER4BB499 - 555273 - 329
18LYSLYSPHEPHE5AA556 - 584330 - 358
28LYSLYSPHEPHE5BB556 - 584330 - 358
19METMETGLUGLU6AA585 - 594359 - 368
29METMETGLUGLU6BB585 - 594359 - 368
110LYSLYSGLNGLN5AA595 - 686369 - 460
210LYSLYSGLNGLN5BB595 - 686369 - 460
111ASPASPLEULEU6AA687 - 696461 - 470
211ASPASPLEULEU6BB687 - 696461 - 470
112LYSLYSSERSER5AA697 - 759471 - 533
212LYSLYSSERSER5BB697 - 759471 - 533
113SERSERHISHIS6AA760 - 823534 - 597
213SERSERHISHIS6BB760 - 823534 - 597
114ILEILEASNASN6AA824 - 837598 - 611
214ILEILEASNASN6BB824 - 837598 - 611
115PROPROTYRTYR5AA838 - 881612 - 655
215PROPROTYRTYR5BB838 - 881612 - 655

NCS oper: (Code: given
Matrix: (0.67739, 0.08674, 0.7305), (0.71762, -0.29631, -0.63026), (0.16179, 0.95114, -0.26297)
Vector: -45.36862, 105.17695, -35.30404)

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Components

#1: Protein ACTIN-LIKE PROTEIN ARP8


Mass: 74813.469 Da / Num. of mol.: 2 / Fragment: C-TERMINAL DOMAIN, RESIDUES 248-881
Source method: isolated from a genetically manipulated source
Details: NUCLEOTIDE-FREE STATE OF THE PROTEIN
Source: (gene. exp.) SACCHAROMYCES CEREVISIAE (brewer's yeast)
Strain: W303-1A / Plasmid: PET28A / Production host: ESCHERICHIA COLI B (bacteria) / Strain (production host): B834 / Variant (production host): PLYSS / References: UniProt: Q12386
#2: Chemical ChemComp-SO4 / SULFATE ION


Mass: 96.063 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: SO4
#3: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 32 / Source method: isolated from a natural source / Formula: H2O
Nonpolymer detailsSULFATE ION (SO4): TWO SULPHATE IONS ARE LOCATED IN THE NUCLEOTIDE-BINDING SITE OF CHAIN A
Sequence detailsTHE CRYSTALLISED PROTEIN CONTAINS THE ARP8 C-TERMINAL DOMAIN FROM RESIDUE 248 TO 881 AND A N- ...THE CRYSTALLISED PROTEIN CONTAINS THE ARP8 C-TERMINAL DOMAIN FROM RESIDUE 248 TO 881 AND A N-TERMINAL EXPRESSION- TAG, COMPRISING RESIDUES MGSSHHHHHHSSGLVPRGSHM

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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: 2.8 Å3/Da / Density % sol: 57 % / Description: NONE
Crystal growpH: 6.5
Details: 18% W/V PEG 5000 MME, 0.28 M LI2SO4, 0.1 M NA-CACODYLATE PH 6.5

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: Diamond / Beamline: I04 / Wavelength: 0.9763
DetectorType: ADSC CCD / Detector: CCD / Date: Mar 12, 2010 / Details: MIRRORS
RadiationMonochromator: DOUBLE CRYSTAL / Protocol: MAD / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9763 Å / Relative weight: 1
ReflectionResolution: 2.7→29 Å / Num. obs: 44062 / % possible obs: 99 % / Observed criterion σ(I): 2 / Redundancy: 4.7 % / Biso Wilson estimate: 83.265 Å2 / Rmerge(I) obs: 0.09 / Net I/σ(I): 20.7
Reflection shellResolution: 2.7→2.85 Å / Redundancy: 3.5 % / Rmerge(I) obs: 0.35 / Mean I/σ(I) obs: 3.5 / % possible all: 96.9

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Processing

Software
NameVersionClassification
REFMAC5.5.0088refinement
MOSFLMdata reduction
SCALAdata scaling
SHARPphasing
RefinementMethod to determine structure: MAD
Starting model: NONE

Resolution: 2.7→28.89 Å / Cor.coef. Fo:Fc: 0.938 / Cor.coef. Fo:Fc free: 0.892 / SU B: 34.253 / SU ML: 0.322 / Cross valid method: THROUGHOUT / ESU R: 1.649 / ESU R Free: 0.388 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. N-TERMINAL TAG AND RESIDUES 248 TO 258 OF ARP8 HAVE NOT BEEN LOCATED IN THE ELECTRON DENSITY MAP.
RfactorNum. reflection% reflectionSelection details
Rfree0.28642 2190 5 %RANDOM
Rwork0.2228 ---
obs0.226 41860 98.92 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso mean: 86.485 Å2
Baniso -1Baniso -2Baniso -3
1--1.06 Å20 Å20.44 Å2
2--2.53 Å20 Å2
3----1.09 Å2
Refinement stepCycle: LAST / Resolution: 2.7→28.89 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms10072 0 10 32 10114
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.0170.02210306
X-RAY DIFFRACTIONr_bond_other_d
X-RAY DIFFRACTIONr_angle_refined_deg1.8411.95813980
X-RAY DIFFRACTIONr_angle_other_deg
X-RAY DIFFRACTIONr_dihedral_angle_1_deg7.88151244
X-RAY DIFFRACTIONr_dihedral_angle_2_deg40.94425.412510
X-RAY DIFFRACTIONr_dihedral_angle_3_deg23.675151838
X-RAY DIFFRACTIONr_dihedral_angle_4_deg23.1281542
X-RAY DIFFRACTIONr_chiral_restr0.1330.21558
X-RAY DIFFRACTIONr_gen_planes_refined0.0080.0217818
X-RAY DIFFRACTIONr_gen_planes_other
X-RAY DIFFRACTIONr_nbd_refined
X-RAY DIFFRACTIONr_nbd_other
X-RAY DIFFRACTIONr_nbtor_refined
X-RAY DIFFRACTIONr_nbtor_other
X-RAY DIFFRACTIONr_xyhbond_nbd_refined
X-RAY DIFFRACTIONr_xyhbond_nbd_other
X-RAY DIFFRACTIONr_metal_ion_refined
X-RAY DIFFRACTIONr_metal_ion_other
X-RAY DIFFRACTIONr_symmetry_vdw_refined
X-RAY DIFFRACTIONr_symmetry_vdw_other
X-RAY DIFFRACTIONr_symmetry_hbond_refined
X-RAY DIFFRACTIONr_symmetry_hbond_other
X-RAY DIFFRACTIONr_symmetry_metal_ion_refined
X-RAY DIFFRACTIONr_symmetry_metal_ion_other
X-RAY DIFFRACTIONr_mcbond_it0.7631.56250
X-RAY DIFFRACTIONr_mcbond_other
X-RAY DIFFRACTIONr_mcangle_it1.433210196
X-RAY DIFFRACTIONr_mcangle_other
X-RAY DIFFRACTIONr_scbond_it2.01234056
X-RAY DIFFRACTIONr_scbond_other
X-RAY DIFFRACTIONr_scangle_it3.2574.53784
X-RAY DIFFRACTIONr_scangle_other
X-RAY DIFFRACTIONr_long_range_B_refined
X-RAY DIFFRACTIONr_long_range_B_other
X-RAY DIFFRACTIONr_rigid_bond_restr
X-RAY DIFFRACTIONr_sphericity_free
X-RAY DIFFRACTIONr_sphericity_bonded
Refine LS restraints NCS

Ens-ID: 1 / Refine-ID: X-RAY DIFFRACTION

Dom-IDAuth asym-IDNumberTypeRms dev position (Å)Weight position
1A2173medium positional0.410.5
2B2173medium positional0.410.5
1A2771loose positional1.375
2B2771loose positional1.375
1A2173medium thermal3.372
2B2173medium thermal3.372
1A2771loose thermal3.4810
2B2771loose thermal3.4810
LS refinement shellResolution: 2.701→2.771 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.396 148 -
Rwork0.424 2901 -
obs--93.76 %
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)
13.381-0.97790.43673.2985-0.06473.01940.2323-0.3620.04270.1002-0.45690.4481-0.002-0.32520.22470.0908-0.06590.04170.1971-0.18060.315-116.28777.5903-9.1342
23.4204-0.2073-0.29354.3455-1.58444.20990.10880.5457-0.2514-0.30150.19630.30640.1262-0.443-0.3050.21230.1953-0.06620.3717-0.0510.1431-129.994424.9404-44.4944
35.36750.92081.70367.6145-0.10210.20020.34170.78460.0019-0.6138-0.46020.02510.10520.66430.11850.17820.1665-0.06580.2376-0.08870.1884-113.6663.5589-33.657
49.2484-3.6283-2.49935.54531.9523.08370.01580.63910.67120.11480.38150.1349-0.4944-0.7177-0.39730.44650.45370.14220.68930.25410.318-145.734444.3656-40.4394
54.3395-1.13330.22732.21840.42622.30690.1326-0.5931-0.26590.2236-0.27120.08720.2020.14240.13860.1762-0.07770.01650.28560.15510.2527-93.8361-4.0985-1.4436
63.6319-0.1864-1.75564.7258-1.04844.44350.0940.7008-0.0164-0.4284-0.0394-0.4788-0.25280.1499-0.05460.34180.15140.10970.44280.01110.1903-110.341739.6253-54.3951
75.62050.48990.9414.19350.19863.77630.47810.5026-0.6915-0.6317-0.56470.01820.33760.38610.08660.39320.2623-0.06390.2051-0.04380.3238-96.9339-14.095-26.5622
87.29460.9649-3.37641.368-1.65594.64010.52991.38851.0685-0.26930.27890.3454-0.9015-0.9547-0.80880.92140.49680.22940.77690.52340.4856-131.85256.8699-57.8972
99.1919-0.284.62322.9117-1.20236.56760.74380.5774-0.9361-0.8492-0.8481-0.08911.04290.68530.10420.59510.39880.0480.3327-0.04520.3378-95.5146-14.4341-28.0633
109.7513.2733-3.10842.6487-0.58473.0103-0.02380.81660.2521-0.17880.31240.1581-0.685-0.5909-0.28861.04730.47510.27340.66930.46150.6885-131.957858.8037-57.3269
111.6252-2.26940.13493.60210.50672.04090.24150.37660.7152-0.3466-0.6039-0.7611-0.44480.5150.36250.3051-0.2410.09060.52750.11540.5945-89.686813.8201-7.8889
124.5661.0404-3.76772.2305-1.78123.5960.3701-0.92650.52490.9589-0.1445-0.4302-0.82710.8103-0.22560.7664-0.0007-0.10420.5775-0.10710.3828-110.793541.4331-34.6484
1310.51121.2456-0.8146.60312.03996.280.2585-0.63170.90581.17980.0473-0.5031-0.13930.8569-0.30590.3274-0.1856-0.20060.69790.04580.3503-74.68269.133716.609
142.99343.5040.17456.78163.48124.48610.1638-0.3373-0.99980.60260.0758-1.16450.21351.3994-0.23960.4624-0.0148-0.0521.49310.42941.1371-83.213138.9266-41.9368
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A259 - 293
2X-RAY DIFFRACTION1A403 - 494
3X-RAY DIFFRACTION1A836 - 881
4X-RAY DIFFRACTION2B259 - 293
5X-RAY DIFFRACTION2B403 - 494
6X-RAY DIFFRACTION2B836 - 881
7X-RAY DIFFRACTION3A294 - 305
8X-RAY DIFFRACTION3A381 - 402
9X-RAY DIFFRACTION4B294 - 305
10X-RAY DIFFRACTION4B381 - 402
11X-RAY DIFFRACTION5A495 - 529
12X-RAY DIFFRACTION5A700 - 759
13X-RAY DIFFRACTION5A824 - 835
14X-RAY DIFFRACTION6B495 - 529
15X-RAY DIFFRACTION6B700 - 759
16X-RAY DIFFRACTION6B824 - 835
17X-RAY DIFFRACTION7A530 - 619
18X-RAY DIFFRACTION8B530 - 619
19X-RAY DIFFRACTION9A306 - 380
20X-RAY DIFFRACTION10B306 - 380
21X-RAY DIFFRACTION11A620 - 699
22X-RAY DIFFRACTION12B620 - 699
23X-RAY DIFFRACTION13A760 - 823
24X-RAY DIFFRACTION14B760 - 823

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