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Solution structure of Staufen1 dsRBD3+4 - hARF1 SBS dsRNA complex.
Authors
Yadav, D.K., Lukavsky, P.J.
Assembly
Double-stranded RNA-binding protein Staufen homolog 1/RNA Complex
Entity
1. Double-stranded RNA-binding protein Staufen homolog 1/RNA Complex, entity 1 (polymer, Thiol state: not present), 177 monomers, 19732.80 Da Detail

GSHMKSEISQ VFEIALKRNL PVNFEVARES GPPHMKNFVT KVSVGEFVGE GEGKSKKISK KNAAIAVLEE LKKLPPLPAV ERVKPRIKKK TKPIVKPQTS PEYGQGINPI SRLAQIQQAK KEKEPEYTLL TERGLPRRRE FVMQVKVGNH TAEGTGTNKK VAKRNAAENM LEILGFK


2. Double-stranded RNA-binding protein Staufen homolog 1/RNA Complex, entity 2 (polymer, Thiol state: not present), 34 monomers, 10962.45 Da Detail

GGCAGAAGCU GCCUCUUCGG AGGCAGUUUC UGCC


Total weight
30695.25 Da
Max. entity weight
19732.8 Da
Source organism
Homo sapiens
Exptl. method
solution NMR
Refine. method
simulated annealingna
Data set
assigned_chemical_shifts
Chem. Shift Complete
Sequence coverage: 90.5 %, Completeness: 84.2 %, Completeness (bb): 89.5 % Detail

Polymer type: polypeptide(L) polyribonucleotide

Total1H13C15N
All84.2 % (2265 of 2690)85.1 % (1228 of 1443)83.9 % (879 of 1048)79.4 % (158 of 199)
Backbone89.5 % (1262 of 1410)90.7 % (506 of 558)89.0 % (612 of 688)87.8 % (144 of 164)
Sidechain79.5 % (1148 of 1444)81.6 % (722 of 885)78.6 % (412 of 524)40.0 % (14 of 35)
Aromatic59.6 % (143 of 240)80.8 % (97 of 120)45.5 % (46 of 101) 0.0 % (0 of 19)
Methyl91.0 % (171 of 188)90.4 % (85 of 94)91.5 % (86 of 94)

1. entity 1

GSHMKSEISQ VFEIALKRNL PVNFEVARES GPPHMKNFVT KVSVGEFVGE GEGKSKKISK KNAAIAVLEE LKKLPPLPAV ERVKPRIKKK TKPIVKPQTS PEYGQGINPI SRLAQIQQAK KEKEPEYTLL TERGLPRRRE FVMQVKVGNH TAEGTGTNKK VAKRNAAENM LEILGFK

2. entity 2

GGCAGAAGCU GCCUCUUCGG AGGCAGUUUC UGCC

Sample #1

Solvent system 90-95% H2O/5-10% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 310 uM [U-15N] Staufen1 dsRBD4, 310 uM Unlabelled hARF1, 90-95% H2O/5-10% D2O


#NameIsotope labelingTypeConcentration
1Staufen1 dsRBD4[U-15N]310 uM
2hARF1Unlabelled310 uM
Sample #2

Solvent system 90-95% H2O/5-10% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 405 uM [U-13C; U-15N] Staufen1 dsRBD4, 405 uM Unlabelled hARF1, 90-95% H2O/5-10% D2O


#NameIsotope labelingTypeConcentration
3Staufen1 dsRBD4[U-13C; U-15N]405 uM
4hARF1Unlabelled405 uM
Sample #3

Solvent system 90-95% H2O/5-10% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 310 uM Unlabelled hARF1, 90-95% H2O/5-10% D2O


#NameIsotope labelingTypeConcentration
5hARF1Unlabelled310 uM
Sample #4

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 478 uM [U-13C] hARF1, 100% D2O


#NameIsotope labelingTypeConcentration
6hARF1[U-13C]478 uM
Sample #5

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 354 uM [U-15N] Staufen1 dsRBD4, 354 uM [U-13C] hARF1, 100% D2O


#NameIsotope labelingTypeConcentration
7Staufen1 dsRBD4[U-15N]354 uM
8hARF1[U-13C]354 uM
Sample #6

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 451 uM [U-13C; U-15N] Staufen1 dsRDB4, 451 uM Unlabelled hARF1, 100% D2O


#NameIsotope labelingTypeConcentration
9Staufen1 dsRDB4[U-13C; U-15N]451 uM
10hARF1Unlabelled451 uM
Sample #7

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 310 uM Unlabelled hARF1, 100% D2O


#NameIsotope labelingTypeConcentration
11hARF1Unlabelled310 uM
Sample #8

Solvent system 90-95% H2O/5-10% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 417 uM [U-15N] Staufen1 dsRBD3, 417 uM Unlabelled hARF1, 90-95% H2O/5-10% D2O


#NameIsotope labelingTypeConcentration
12Staufen1 dsRBD3[U-15N]417 uM
13hARF1Unlabelled417 uM
Sample #9

Solvent system 90-95% H2O/5-10% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 458 uM [U-13C; U-15N] Staufen1 dsRBD3, 458 uM Unlabelled hARF1, 90-95% H2O/5-10% D2O


#NameIsotope labelingTypeConcentration
14hARF1Unlabelled458 uM
15Staufen1 dsRBD3[U-13C; U-15N]458 uM
Sample #10

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 458 uM [U-13C; U-15N] Staufen1 dsRBD3, 458 uM Unlabelled hARF1, 100% D2O


#NameIsotope labelingTypeConcentration
16hARF1Unlabelled458 uM
17Staufen1 dsRBD3[U-13C; U-15N]458 uM
Sample #11

Solvent system 100% D2O, Pressure 1 atm, Temperature 308 K, pH 6.5, Details 157 uM [U-13C; U-15N] Staufen1 dsRBD3, 157 uM Unlabelled Staufen1 dsRBD4, 157 uM Unlabelled hARF1, 100% D2O


#NameIsotope labelingTypeConcentration
18Staufen1 dsRBD4Unlabelled157 uM
19hARF1Unlabelled157 uM
20Staufen1 dsRBD3[U-13C; U-15N]157 uM

LACS Plot; CA
Referencing offset: -0.07 ppm, Outliers: 1 Detail
LACS Plot; CB
Referencing offset: -0.07 ppm, Outliers: 1 Detail
LACS Plot; HA
Referencing offset: -0.11 ppm, Outliers: 1 Detail
LACS Plot; CO
Referencing offset: 0.15 ppm, Outliers: 1 Detail
Protein Blocks Logo
Calculated from 20 models in PDB: 6SDW, Strand ID: A Detail


Release date
2020-01-06
Citation
Staufen1 reads out structure and sequence features in ARF1 dsRNA for target recognition
Yadav, D.K., Zigackova, D., Zlobina, M., Klumpler, T., Beaumont, C., Kubickova, M., Vanacova, S., Lukavsky, P.J.
Nucleic Acids Res. (2019), 48, 2091-2106, PubMed 31875226 , DOI 10.1093/nar/gkz1163 ,
Related entities 1. Double-stranded RNA-binding protein Staufen homolog 1/RNA Complex, entity 1, : 1 : 2 : 122 entities Detail
Experiments performed 59 experiments Detail
Chemical shift validation 4 contents Detail
Keywords NMR structure of RNA-protein complex, RNA binding domain., RNA binding protein, Staufen mediated mRNA decay, Staufen1 protein, dsRBD-dsRNA binding protein