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- PDB-3hm9: Crystal structure of T. thermophilus Argonaute complexed with DNA... -

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Basic information

Entry
Database: PDB / ID: 3hm9
TitleCrystal structure of T. thermophilus Argonaute complexed with DNA guide strand and 19-nt RNA target strand
Components
  • 5'-D(P*TP*GP*AP*GP*GP*TP*AP*GP*TP*AP*GP*GP*TP*TP*GP*TP*AP*TP*AP*GP*T)-3'
  • 5'-R(*UP*AP*UP*AP*CP*AP*AP*CP*CP*UP*AP*CP*UP*AP*CP*CP*UP*CP*G)-3'
  • Argonaute
KeywordsNucleic Acid Binding Protein/DNA/RNA / argonaute / protein-DNA-RNA complex / Nucleic Acid Binding Protein-DNA-RNA COMPLEX
Function / homology
Function and homology information


Hydrolases; Acting on ester bonds; Site specific endodeoxyribonucleases: cleavage is not sequence specific (deleted sub-subclass) / clearance of foreign intracellular DNA / DNA endonuclease activity / manganese ion binding / DNA replication / DNA binding / RNA binding
Similarity search - Function
paz domain - #50 / Alpha-D-Glucose-1,6-Bisphosphate; Chain A, domain 4 - #60 / Alpha-Beta Plaits - #2620 / Argonaute, PAZ domain / Argonaute PAZ domain / paz domain / Piwi domain / Piwi domain profile. / Piwi domain / Piwi ...paz domain - #50 / Alpha-D-Glucose-1,6-Bisphosphate; Chain A, domain 4 - #60 / Alpha-Beta Plaits - #2620 / Argonaute, PAZ domain / Argonaute PAZ domain / paz domain / Piwi domain / Piwi domain profile. / Piwi domain / Piwi / PAZ domain profile. / PAZ domain / Alpha-D-Glucose-1,6-Bisphosphate; Chain A, domain 4 / Response regulator / Beta Complex / Ribonuclease H-like superfamily/Ribonuclease H / Nucleotidyltransferase; domain 5 / Ribonuclease H superfamily / Ribonuclease H-like superfamily / Alpha-Beta Plaits / Rossmann fold / 2-Layer Sandwich / 3-Layer(aba) Sandwich / Mainly Beta / Alpha Beta
Similarity search - Domain/homology
DNA / DNA (> 10) / RNA / RNA (> 10) / Protein argonaute
Similarity search - Component
Biological speciesThermus thermophilus (bacteria)
MethodX-RAY DIFFRACTION / SYNCHROTRON / MOLECULAR REPLACEMENT / Resolution: 3.3 Å
AuthorsWang, Y. / Li, H. / Sheng, G. / Patel, D.J.
CitationJournal: Nature / Year: 2009
Title: Nucleation, propagation and cleavage of target RNAs in Ago silencing complexes.
Authors: Wang, Y. / Juranek, S. / Li, H. / Sheng, G. / Wardle, G.S. / Tuschl, T. / Patel, D.J.
History
DepositionMay 28, 2009Deposition site: RCSB / Processing site: RCSB
Revision 1.0Oct 6, 2009Provider: repository / Type: Initial release
Revision 1.1Jul 13, 2011Group: Advisory / Refinement description ...Advisory / Refinement description / Source and taxonomy / Version format compliance
Revision 1.2Feb 21, 2024Group: Data collection / Database references / Derived calculations
Category: chem_comp_atom / chem_comp_bond ...chem_comp_atom / chem_comp_bond / database_2 / struct_conn / struct_site
Item: _database_2.pdbx_DOI / _database_2.pdbx_database_accession ..._database_2.pdbx_DOI / _database_2.pdbx_database_accession / _struct_conn.ptnr1_auth_asym_id / _struct_conn.ptnr1_auth_comp_id / _struct_conn.ptnr1_auth_seq_id / _struct_conn.ptnr1_label_asym_id / _struct_conn.ptnr1_label_atom_id / _struct_conn.ptnr1_label_comp_id / _struct_conn.ptnr1_label_seq_id / _struct_conn.ptnr2_auth_asym_id / _struct_conn.ptnr2_auth_comp_id / _struct_conn.ptnr2_auth_seq_id / _struct_conn.ptnr2_label_asym_id / _struct_conn.ptnr2_label_atom_id / _struct_conn.ptnr2_label_comp_id / _struct_conn.ptnr2_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: Argonaute
X: 5'-D(P*TP*GP*AP*GP*GP*TP*AP*GP*TP*AP*GP*GP*TP*TP*GP*TP*AP*TP*AP*GP*T)-3'
Y: 5'-R(*UP*AP*UP*AP*CP*AP*AP*CP*CP*UP*AP*CP*UP*AP*CP*CP*UP*CP*G)-3'
hetero molecules


Theoretical massNumber of molelcules
Total (without water)89,3085
Polymers89,2603
Non-polymers492
Water00
1


  • Idetical with deposited unit
  • defined by software
TypeNameSymmetry operationNumber
identity operation1_555x,y,z1
Buried area5190 Å2
ΔGint-62 kcal/mol
Surface area32990 Å2
MethodPISA
Unit cell
Length a, b, c (Å)111.916, 111.916, 175.551
Angle α, β, γ (deg.)90.000, 90.000, 90.000
Int Tables number96
Space group name H-MP43212

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Components

#1: Protein Argonaute


Mass: 76728.734 Da / Num. of mol.: 1
Source method: isolated from a genetically manipulated source
Source: (gene. exp.) Thermus thermophilus (bacteria) / Strain: HB27 / Gene: TT_P0026 / Plasmid: PET-Sumo / Production host: Escherichia coli (E. coli) / Strain (production host): ROSETTA 2 (DE3)pLysS / References: UniProt: Q746M7
#2: DNA chain 5'-D(P*TP*GP*AP*GP*GP*TP*AP*GP*TP*AP*GP*GP*TP*TP*GP*TP*AP*TP*AP*GP*T)-3'


Mass: 6588.266 Da / Num. of mol.: 1 / Source method: obtained synthetically
#3: RNA chain 5'-R(*UP*AP*UP*AP*CP*AP*AP*CP*CP*UP*AP*CP*UP*AP*CP*CP*UP*CP*G)-3'


Mass: 5942.584 Da / Num. of mol.: 1 / Source method: obtained synthetically
#4: Chemical ChemComp-MG / MAGNESIUM ION


Mass: 24.305 Da / Num. of mol.: 2 / Source method: obtained synthetically / Formula: Mg

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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: 3.08 Å3/Da / Density % sol: 60.05 %
Crystal growTemperature: 308 K / Method: vapor diffusion, hanging drop / pH: 7
Details: 50 mM MgCl2, 1.0 M Na tartrate, 50 mM Tris.HCl pH 7.0, VAPOR DIFFUSION, HANGING DROP, temperature 308 K

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Data collection

DiffractionMean temperature: 100 K
Diffraction sourceSource: SYNCHROTRON / Site: APS / Beamline: 24-ID-C / Wavelength: 0.979 Å
DetectorType: MAR scanner 300 mm plate / Detector: IMAGE PLATE / Date: Mar 17, 2009
RadiationProtocol: SINGLE WAVELENGTH / Monochromatic (M) / Laue (L): M / Scattering type: x-ray
Radiation wavelengthWavelength: 0.979 Å / Relative weight: 1
ReflectionResolution: 3.3→50 Å / Num. obs: 17434 / % possible obs: 99.5 % / Observed criterion σ(F): -1 / Redundancy: 7.3 % / Rmerge(I) obs: 0.06 / Net I/σ(I): 26.3
Reflection shellResolution: 3.3→3.38 Å / Redundancy: 6.8 % / Rmerge(I) obs: 0.444 / Mean I/σ(I) obs: 2.6 / Num. unique all: 1128 / % possible all: 100

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Processing

Software
NameVersionClassificationNB
REFMAC5.5.0088refinement
PDB_EXTRACT3.005data extraction
HKL-2000data collection
HKL-2000data reduction
HKL-2000data scaling
PHASERphasing
RefinementMethod to determine structure: MOLECULAR REPLACEMENT / Resolution: 3.3→20 Å / Cor.coef. Fo:Fc: 0.93 / Cor.coef. Fo:Fc free: 0.879 / Occupancy max: 1 / Occupancy min: 1 / SU B: 60.16 / SU ML: 0.466 / TLS residual ADP flag: LIKELY RESIDUAL / Cross valid method: THROUGHOUT / σ(F): 0 / ESU R Free: 0.549 / Stereochemistry target values: MAXIMUM LIKELIHOOD
Details: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS U VALUES : RESIDUAL ONLY
RfactorNum. reflection% reflectionSelection details
Rfree0.289 871 5 %RANDOM
Rwork0.237 ---
obs0.239 17255 99.62 %-
Solvent computationIon probe radii: 0.8 Å / Shrinkage radii: 0.8 Å / VDW probe radii: 1.4 Å / Solvent model: MASK
Displacement parametersBiso max: 77.24 Å2 / Biso mean: 58.365 Å2 / Biso min: 20 Å2
Baniso -1Baniso -2Baniso -3
1-0.17 Å20 Å20 Å2
2--0.17 Å20 Å2
3----0.35 Å2
Refinement stepCycle: LAST / Resolution: 3.3→20 Å
ProteinNucleic acidLigandSolventTotal
Num. atoms4631 606 2 0 5239
Refine LS restraints
Refine-IDTypeDev idealDev ideal targetNumber
X-RAY DIFFRACTIONr_bond_refined_d0.010.0225421
X-RAY DIFFRACTIONr_angle_refined_deg1.3982.1057510
X-RAY DIFFRACTIONr_dihedral_angle_1_deg7.185649
X-RAY DIFFRACTIONr_dihedral_angle_2_deg34.40721.294170
X-RAY DIFFRACTIONr_dihedral_angle_3_deg22.48415627
X-RAY DIFFRACTIONr_dihedral_angle_4_deg18.3271547
X-RAY DIFFRACTIONr_chiral_restr0.0850.2866
X-RAY DIFFRACTIONr_gen_planes_refined0.0050.0213970
X-RAY DIFFRACTIONr_mcbond_it0.321.53247
X-RAY DIFFRACTIONr_mcangle_it0.64525003
X-RAY DIFFRACTIONr_scbond_it1.03632174
X-RAY DIFFRACTIONr_scangle_it1.7464.52507
LS refinement shellResolution: 3.3→3.383 Å / Total num. of bins used: 20
RfactorNum. reflection% reflection
Rfree0.395 65 -
Rwork0.281 1161 -
all-1226 -
obs--100 %
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)
12.1654-2.0858-0.64444.63750.62220.2793-0.5221-0.72310.91531.29810.6524-0.21570.17950.4573-0.13030.97570.123-0.23531.3049-0.06070.9642-27.508736.353515.7265
21.6027-1.0922-2.2334.752-3.678910.708-0.34410.1211-0.03030.08270.43950.72150.6467-1.2636-0.09540.28310.1028-0.03651.0326-0.07490.8915-34.518718.472712.2599
31.43851.2719-1.18481.2137-1.02321.00470.03371.17450.31040.65110.58970.3991-0.0478-0.9217-0.62332.2491-0.33060.42482.7312-0.38691.999-47.448413.470539.6774
43.6551-2.35471.1823.6171-1.74631.53920.03880.5608-0.1284-0.66440.0580.54770.7735-0.2409-0.09680.6983-0.09540.02241.0234-0.07070.4931-21.52849.96684.9847
51.84891.6143-0.28644.5555-1.58153.3111-0.17650.3695-0.4227-0.35180.1241-0.33370.6434-0.00770.05240.35090.15490.12190.2596-0.06350.4527-8.2283-9.501432.1802
61.47610.5358-0.53962.9772-0.39674.5856-0.0270.599-0.1923-0.34840.047-0.12830.08130.0097-0.01990.2090.14050.13750.3678-0.00090.2884-4.9888.428421.1011
76.7644-2.73851.18893.3307-3.41164.2813-0.39480.48160.5282-0.13490.7520.40120.3979-1.2688-0.35720.505-0.0078-0.02340.69310.05720.7606-23.8775-0.197521.0311
817.1831-2.2448-0.07630.78451.1442.7196-0.20.38841.4018-0.0657-0.33670.1577-0.1795-0.7040.53670.68670.23460.03580.7543-0.00650.9597-22.225721.245929.9138
912.64020.1798-10.35222.5595.627921.6588-0.51661.23051.3857-0.69070.03230.8447-1.183-0.68830.48440.5028-0.0869-0.17560.8734-0.07670.8128-20.139118.118523.733
106.278-2.77483.24575.42754.577410.72560.43130.1867-0.2194-0.1093-0.76660.68820.643-1.33760.33520.8916-0.3046-0.01420.95620.13510.6261-27.8356-1.740720.6488
Refinement TLS group
IDRefine-IDRefine TLS-IDAuth asym-IDAuth seq-ID
1X-RAY DIFFRACTION1A3 - 135
2X-RAY DIFFRACTION2A136 - 180
3X-RAY DIFFRACTION3A181 - 262
4X-RAY DIFFRACTION4A263 - 330
5X-RAY DIFFRACTION5A331 - 521
6X-RAY DIFFRACTION6A522 - 685
7X-RAY DIFFRACTION7X1 - 9
8X-RAY DIFFRACTION8X10 - 16
9X-RAY DIFFRACTION9Y6 - 10
10X-RAY DIFFRACTION10Y11 - 18

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