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- PDB-4x8w: dsRBD3 of Loquacious -

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Basic information

Entry
Database: PDB / ID: 4x8w
TitledsRBD3 of Loquacious
Components
  • Loquacious
  • Loquacious, isoform B
KeywordsGENE REGULATION / double stranded RNA binding domain / dimerization / dicer binding / miRNA processing
Function / homology
Function and homology information


regulatory ncRNA processing / MicroRNA (miRNA) biogenesis / Small interfering RNA (siRNA) biogenesis / female germ-line stem cell asymmetric division / PKR-mediated signaling / regulation of regulatory ncRNA processing / dosage compensation by hyperactivation of X chromosome / RISC complex binding / germ-line stem cell population maintenance / RISC-loading complex ...regulatory ncRNA processing / MicroRNA (miRNA) biogenesis / Small interfering RNA (siRNA) biogenesis / female germ-line stem cell asymmetric division / PKR-mediated signaling / regulation of regulatory ncRNA processing / dosage compensation by hyperactivation of X chromosome / RISC complex binding / germ-line stem cell population maintenance / RISC-loading complex / miRNA metabolic process / RISC complex assembly / siRNA binding / pre-miRNA processing / siRNA processing / RISC complex / central nervous system development / double-stranded RNA binding / nucleus / cytoplasm / cytosol
Similarity search - Function
Double Stranded RNA Binding Domain - #20 / Double-stranded RNA binding motif / Double-stranded RNA binding motif / Double stranded RNA-binding domain (dsRBD) profile. / Double-stranded RNA-binding domain / Double Stranded RNA Binding Domain / 2-Layer Sandwich / Alpha Beta
Similarity search - Domain/homology
Biological speciesDrosophila melanogaster (fruit fly)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MAD / Resolution: 2.647 Å
AuthorsJakob, L. / Treiber, N. / Treiber, T. / Stotz, M. / Meister, G.
CitationJournal: To Be Published
Title: Structure of the dsRBD3 of Loquacious reveals features for dimerization and Dicer1-interaction
Authors: Jakob, L. / Treiber, T. / Treiber, N. / Stotz, M. / Meister, G.
History
DepositionDec 10, 2014Deposition site: RCSB / Processing site: PDBE
Revision 1.0Jan 13, 2016Provider: repository / Type: Initial release

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Structure visualization

Structure viewerMolecule:
MolmilJmol/JSmol

Downloads & links

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Assembly

Deposited unit
A: Loquacious, isoform B
B: Loquacious, isoform B
C: Loquacious, isoform B
D: Loquacious, isoform B
E: Loquacious, isoform B
F: Loquacious, isoform B
G: Loquacious, isoform B
H: Loquacious
hetero molecules


Theoretical massNumber of molelcules
Total (without water)60,7489
Polymers60,6568
Non-polymers921
Water1,20767
1
A: Loquacious, isoform B
B: Loquacious, isoform B
D: Loquacious, isoform B
E: Loquacious, isoform B
hetero molecules


Theoretical massNumber of molelcules
Total (without water)32,7935
Polymers32,7014
Non-polymers921
Water724
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area7980 Å2
ΔGint-64 kcal/mol
Surface area15140 Å2
MethodPISA
2
C: Loquacious, isoform B
F: Loquacious, isoform B
G: Loquacious, isoform B


Theoretical massNumber of molelcules
Total (without water)24,5263
Polymers24,5263
Non-polymers00
Water543
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area3770 Å2
ΔGint-29 kcal/mol
Surface area8750 Å2
MethodPISA
3
H: Loquacious


Theoretical massNumber of molelcules
Total (without water)3,4301
Polymers3,4301
Non-polymers00
Water181
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area0 Å2
ΔGint0 kcal/mol
Surface area4280 Å2
MethodPISA
Unit cell
Length a, b, c (Å)66.018, 112.970, 114.049
Angle α, β, γ (deg.)90.00, 90.00, 90.00
Int Tables number19
Space group name H-MP212121

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Components

#1: Protein
Loquacious, isoform B / R3D1-L / RE14437p


Mass: 8175.219 Da / Num. of mol.: 7 / Fragment: UNP RESIDUES 392-463
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Drosophila melanogaster (fruit fly) / Gene: loqs, CG6866, Dmel_CG6866 / Plasmid: pET32a / Production host: Escherichia coli BL21(DE3) (bacteria) / References: UniProt: Q9VJY9
#2: Protein/peptide Loquacious


Mass: 3429.750 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Details: fragments of loquacious with insufficient electron density
Source: (gene. exp.) Drosophila melanogaster (fruit fly) / Gene: Loquacious / Plasmid: pET32a / Production host: Escherichia coli BL21(DE3) (bacteria)
#3: Chemical ChemComp-GOL / GLYCEROL / GLYCERIN / PROPANE-1,2,3-TRIOL


Mass: 92.094 Da / Num. of mol.: 1 / Source method: obtained synthetically / Formula: C3H8O3
#4: Water ChemComp-HOH / water


Mass: 18.015 Da / Num. of mol.: 67 / Source method: isolated from a natural source / Formula: H2O

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Experimental details

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Experiment

ExperimentMethod: X-RAY DIFFRACTION

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Sample preparation

CrystalDensity Matthews: 3.29 Å3/Da / Density % sol: 62.64 %
Crystal growTemperature: 293 K / Method: vapor diffusion, hanging drop / pH: 7.5
Details: 2% PEG1000 12% PEG8000 50 mM Hepes-NaOH 250 mM NaCl 1mM DTT

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Data collection

DiffractionMean temperature: 80 K
Diffraction sourceSource: SYNCHROTRON / Site: BESSY / Beamline: 14.2 / Wavelength: 0.9184 Å
DetectorType: MARMOSAIC 225 mm CCD / Detector: CCD / Date: Jul 16, 2013
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.9184 Å / Relative weight: 1
ReflectionResolution: 2.647→50 Å / Num. obs: 25522 / % possible obs: 99.9 % / Redundancy: 7.4 % / Rsym value: 0.046 / Net I/σ(I): 33.8
Reflection shellResolution: 2.65→2.81 Å / Redundancy: 7.5 % / Rmerge(I) obs: 0.035 / Mean I/σ(I) obs: 6 / % possible all: 99.9

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Processing

Software
NameVersionClassification
PHENIX(phenix.refine: 1.9_1692)refinement
Cootmodel building
XDSdata reduction
SHARPphasing
XSCALEdata scaling
RefinementMethod to determine structure: MAD / Resolution: 2.647→25.453 Å / SU ML: 0.3 / Cross valid method: FREE R-VALUE / σ(F): 1.34 / Phase error: 26.47 / Stereochemistry target values: ML
RfactorNum. reflection% reflectionSelection details
Rfree0.251 2482 5.18 %Random selection
Rwork0.206 ---
obs0.2084 25451 99.75 %-
Solvent computationShrinkage radii: 0.9 Å / VDW probe radii: 1.11 Å / Solvent model: FLAT BULK SOLVENT MODEL
Refinement stepCycle: LAST / Resolution: 2.647→25.453 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms3626 0 6 67 3699
Refine LS restraints
Refine-IDTypeDev idealNumber
X-RAY DIFFRACTIONf_bond_d0.0093679
X-RAY DIFFRACTIONf_angle_d1.24994
X-RAY DIFFRACTIONf_dihedral_angle_d15.3971265
X-RAY DIFFRACTIONf_chiral_restr0.054590
X-RAY DIFFRACTIONf_plane_restr0.006643
LS refinement shell
Resolution (Å)Rfactor RfreeNum. reflection RfreeRfactor RworkNum. reflection RworkRefine-ID% reflection obs (%)
2.6468-2.69760.3073990.26362534X-RAY DIFFRACTION99
2.6976-2.75260.291380.24952575X-RAY DIFFRACTION100
2.7526-2.81240.30881420.25112485X-RAY DIFFRACTION100
2.8124-2.87780.29471620.24012510X-RAY DIFFRACTION100
2.8778-2.94960.28351240.25022524X-RAY DIFFRACTION100
2.9496-3.02930.28931630.22832520X-RAY DIFFRACTION100
3.0293-3.11820.26991350.21072546X-RAY DIFFRACTION100
3.1182-3.21870.29111410.22792524X-RAY DIFFRACTION100
3.2187-3.33350.27351270.2222537X-RAY DIFFRACTION100
3.3335-3.46670.271500.21072541X-RAY DIFFRACTION100
3.4667-3.6240.2671100.21352529X-RAY DIFFRACTION100
3.624-3.81450.29411490.21162502X-RAY DIFFRACTION100
3.8145-4.05260.24691330.20852538X-RAY DIFFRACTION100
4.0526-4.3640.27641260.18632563X-RAY DIFFRACTION100
4.364-4.80060.22291470.17232485X-RAY DIFFRACTION100
4.8006-5.48910.22031640.18222521X-RAY DIFFRACTION100
5.4891-6.89290.20251600.21462496X-RAY DIFFRACTION100
6.8929-25.45450.19211120.18382463X-RAY DIFFRACTION97
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)
10.47820.0474-0.02190.2952-0.24420.29580.38-0.20710.48820.1291-0.4379-0.37-0.29140.1627-0.250.4498-0.09290.05360.31370.00250.6888.600636.857244.9117
20.0021-0.0082-0.00170.08340.05720.0466-0.0365-0.0292-0.0311-0.4016-0.18210.4173-0.01490.2689-0.00560.364-0.00670.18110.44780.05781.0548-3.821733.46935.8666
30.17680.0769-0.06670.181-0.22960.4673-0.20910.07640.021-0.32980.07580.10670.15440.1875-0.24450.4424-0.0085-0.08950.33490.09790.497424.530444.527821.1003
40.53410.07770.06191.03290.03980.3341-0.03690.28390.1542-0.43280.0580.6653-0.0109-0.0552-0.07720.3340.0086-0.03650.21280.07740.484819.74145.29621.7291
50.13320.08750.05280.16410.04730.02730.03580.08330.42650.52190.0356-0.1527-0.14990.0701-0.00290.3979-0.06730.12480.3841-0.08680.66513.796830.830842.6722
60.21080.1640.20540.28410.06760.24740.48660.09130.3536-0.0732-0.088-0.1552-0.2530.0430.27510.32250.06240.2460.40790.01560.515946.170724.244827.38
70.04830.1566-0.21770.4962-0.68210.98330.26860.1953-0.0019-0.08470.2472-0.17760.0023-0.35460.24950.217-0.0347-0.02120.4025-0.3254-0.068439.094715.008129.0946
80.01520.0012-0.0240.02240.00690.02410.1504-0.02910.13910.0535-0.12020.0660.0510.0732-0.00010.42460.03720.02520.5129-0.04770.633720.21122.867633.3307
90.30540.1134-0.01420.1345-0.08490.08470.0349-0.0851-0.0185-0.1628-0.0377-0.1945-0.16020.1972-0.09170.32910.01930.02060.3613-0.08490.2137.13415.539934.2065
100.29410.0617-0.11920.20380.1620.23050.21280.08770.236-0.07380.07760.0182-0.23910.08280.75680.37430.00810.1910.2405-0.06080.041431.411822.806133.5381
110.88970.26740.08550.0968-0.05890.67060.34980.01920.68290.03050.0009-0.1118-0.21220.14460.72610.26610.00580.08560.3856-0.15890.392138.909924.592834.2368
120.1092-0.0720.15840.7301-0.05840.2403-0.0718-0.20230.01890.36770.14190.26550.1178-0.55130.12510.5332-0.0380.11240.6839-0.09610.371423.703510.993748.7455
130.158-0.07520.04780.0416-0.05390.25670.1982-0.227-0.03130.00280.1198-0.03860.13440.18040.37890.71190.1756-0.0580.6731-0.2545-0.194235.414811.793450.2413
140.38740.0453-0.0640.1774-0.13170.1032-0.0477-0.53390.26720.29090.057-0.0778-0.21270.1151-0.17490.49460.03670.00520.3615-0.10540.22538.911321.556346.7762
150.0035-0.0075-0.01130.00620.01850.0304-0.0672-0.07960.06180.21710.1211-0.1634-0.05320.16890.24960.39930.088-0.57460.46-0.03690.390839.864952.716134.2965
160.00870.02330.00250.04110.01140.0420.0539-0.1690.06830.0281-0.1051-0.29760.0690.0430.00010.37680.1376-0.07410.41050.04410.569138.232142.744530.2815
170.00050.00220.010.00160.00080.02140.02690.3754-0.1861-0.2393-0.0044-0.1673-0.0050.0160.00030.82410.0579-0.06010.610.02170.474835.044144.629110.3002
180.0121-0.0153-0.04330.27150.05660.02240.0337-0.1783-0.1199-0.23840.0021-0.3740.13380.0110.03140.4880.0174-0.01290.40690.12890.422633.613845.144824.4375
190.0359-0.0510.01820.0822-0.01420.01910.17960.0010.03540.1379-0.0167-0.3084-0.1840.044-0.00010.4272-0.04490.04030.38360.10120.308233.372352.462427.5243
200.0156-0.01590.01770.011-0.01150.01370.20870.0923-0.19780.2054-0.08960.14260.47030.1209-0.00020.3814-0.0469-0.03430.3953-0.16390.3522-6.122213.451133.1118
210.07550.15030.02620.28970.04840.00840.21990.4051-0.00190.1897-0.3278-0.46360.06790.0154-0.00370.37030.07140.01060.4987-0.01910.32494.058415.635935.934
220.4099-0.41660.11110.98620.05140.07280.1394-0.0690.03350.2242-0.1618-0.35790.6749-0.093-0.1440.4196-0.0884-0.02480.446-0.13260.29251.609122.32146.4768
230.4101-0.44470.12680.5098-0.12510.04180.1860.04390.05430.0469-0.57630.27190.1154-0.1732-0.06780.3409-0.05950.00080.4617-0.13530.3707-4.817821.741938.2783
240.13340.07550.04750.03480.02180.00760.2388-0.41160.11070.3191-0.3175-0.08080.2593-0.1783-0.00080.4083-0.0453-0.04050.3521-0.11080.335138.350314.442245.7655
250.70370.2062-0.06960.971-0.15121.7516-0.09020.19640.1816-0.14540.19470.2163-0.0127-0.53950.56010.36380.00850.0670.4732-0.10270.308429.018116.097242.1457
Refinement TLS group
IDRefine-IDRefine TLS-IDSelection details
1X-RAY DIFFRACTION1chain 'E' and (resid 389 through 414 )
2X-RAY DIFFRACTION2chain 'E' and (resid 415 through 431 )
3X-RAY DIFFRACTION3chain 'E' and (resid 432 through 463 )
4X-RAY DIFFRACTION4chain 'D' and (resid 391 through 431 )
5X-RAY DIFFRACTION5chain 'D' and (resid 432 through 463 )
6X-RAY DIFFRACTION6chain 'C' and (resid 391 through 405 )
7X-RAY DIFFRACTION7chain 'C' and (resid 406 through 414 )
8X-RAY DIFFRACTION8chain 'C' and (resid 415 through 422 )
9X-RAY DIFFRACTION9chain 'C' and (resid 423 through 434 )
10X-RAY DIFFRACTION10chain 'C' and (resid 435 through 443 )
11X-RAY DIFFRACTION11chain 'C' and (resid 444 through 462 )
12X-RAY DIFFRACTION12chain 'G' and (resid 390 through 405 )
13X-RAY DIFFRACTION13chain 'G' and (resid 406 through 414 )
14X-RAY DIFFRACTION14chain 'G' and (resid 415 through 433 )
15X-RAY DIFFRACTION15chain 'B' and (resid 392 through 405 )
16X-RAY DIFFRACTION16chain 'B' and (resid 406 through 413 )
17X-RAY DIFFRACTION17chain 'B' and (resid 414 through 422 )
18X-RAY DIFFRACTION18chain 'B' and (resid 423 through 443 )
19X-RAY DIFFRACTION19chain 'B' and (resid 444 through 463 )
20X-RAY DIFFRACTION20chain 'A' and (resid 391 through 404 )
21X-RAY DIFFRACTION21chain 'A' and (resid 405 through 413 )
22X-RAY DIFFRACTION22chain 'A' and (resid 414 through 443 )
23X-RAY DIFFRACTION23chain 'A' and (resid 444 through 462 )
24X-RAY DIFFRACTION24chain 'F' and (resid 429 through 443 )
25X-RAY DIFFRACTION25chain 'F' and (resid 444 through 463 )

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