BMRBj - BMREntryMaster

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BMRB: 34466

6TR0

Solution structure of U2AF2 RRM1,2

Authors

Kang, H.S., Sattler, M.

Assembly

Splicing factor U2AF 65 kDa subunit

Entity

1. Splicing factor U2AF 65 kDa subunit (polymer, Thiol state: all free), 205 monomers, 22322.29 Da Detail

AMVGSQMTRQ ARRLYVGNIP FGITEEAMMD FFNAQMRLGG LTQAPGNPVL AVQINQDKNF AFLEFRSVDE TTQAMAFDGI IFQGQSLKIR RPHDYQPLPG MSENPSVYVP GVVSTVVPDS AHKLFIGGLP NYLNDDQVKE LLTSFGPLKA FNLVKDSATG LSKGYAFCEY VDINVTDQAI AGLNGMQLGD KKLLVQRASV GAKNA

Formula weight

22322.29 Da

Source organism

Exptl. method

Refine. method

simulated annealing

Data set

assigned_chemical_shifts

Chem. Shift Complete

Sequence coverage: 98.5 %, Completeness: 86.7 %, Completeness (bb): 84.4 % Detail

Polymer type: polypeptide(L)

	Total	¹H	¹³C	¹⁵N
All	86.7 % (2046 of 2359)	92.5 % (1131 of 1223)	76.6 % (702 of 916)	96.8 % (213 of 220)
Backbone	84.4 % (1020 of 1208)	96.7 % (404 of 418)	71.5 % (426 of 596)	97.9 % (190 of 194)
Sidechain	90.4 % (1208 of 1337)	90.3 % (727 of 805)	90.5 % (458 of 506)	88.5 % (23 of 26)
Aromatic	80.7 % (142 of 176)	89.8 % (79 of 88)	71.6 % (63 of 88)
Methyl	98.3 % (230 of 234)	97.4 % (114 of 117)	99.1 % (116 of 117)

1. entity 1

AMVGSQMTRQ ARRLYVGNIP FGITEEAMMD FFNAQMRLGG LTQAPGNPVL AVQINQDKNF AFLEFRSVDE TTQAMAFDGI IFQGQSLKIR RPHDYQPLPG MSENPSVYVP GVVSTVVPDS AHKLFIGGLP NYLNDDQVKE LLTSFGPLKA FNLVKDSATG LSKGYAFCEY VDINVTDQAI AGLNGMQLGD KKLLVQRASV GAKNA

Show sample condition

Sample

Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

Protein Blocks Logo

Calculated from 10 models in PDB: 6TR0, Strand ID: A Detail

Release date

2020-04-26

Citation

An autoinhibitory intramolecular interaction proof-reads RNA recognition by the essential splicing factor U2AF2
Kang, H.S., Sanchez-Rico, C., Ebersberger, S., Sutandy, F.X.R., Busch, A., Welte, T., Stehle, R., Hipp, C., Schulz, L., Buchbender, A., Zarnack, K., Konig, J., Sattler, M.
Proc. Natl. Acad. Sci. U. S. A. (2020), 117, 7140-7149, PubMed 32188783 , DOI 10.1073/pnas.1913483117 , Abstract

Related entities 1. Splicing factor U2AF 65 kDa subunit, : 1 : 6 : 194 entities Detail

Interaction partners 1. Splicing factor U2AF 65 kDa subunit, : 97 interactors Detail

More details for 97 interactors identified by 22 citations

Experiments performed 11 experiments Detail

1 3D 1H-15N NOESY

Instrument

Bruker AVANCE III - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

2 3D 1H-13C NOESY

Instrument

Bruker AVANCE III - 800 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

3 3D HCCH-TOCSY

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

4 3D HCCH-COSY

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

5 2D 1H-13C HSQC aromatic

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

6 3D CBCA(CO)NH

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

7 3D HNCACB

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

8 3D HNCO

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

9 3D HNCACO

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

10 2D 1H-15N HSQC

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

11 3D H(CCO)NH

Instrument

Bruker AVANCE III - 600 MHz

Sample

State isotropic, Solvent system 90% H2O/10% D2O, Pressure 1 atm, Temperature 298 K, pH 6.5, Details 0.5 mM [U-13C; U-15N] U2AF2, 90% H2O/10% D2O

#	Name	Isotope labeling	Type	Concentration
1	U2AF2	[U-13C; U-15N]		0.5 mM
2	sodium phosphoate	natural abundance		20 mM
3	sodium chloride	natural abundance		50 mM
4	DTT	natural abundance		2 mM

Chemical shift validation 3 contents Detail

Properties

Input source #1: Assigned chemical shifts - Assigned chemical shifts	bmr34466_3.str
Diamagnetism of the molecular assembly	True (excluding Oxygen atoms)
Whether the assembly has a disulfide bond	None
Whether the assembly has a other bond	None
Whether the assembly contains a cyclic polymer	None
Overall data processing status	Warning

Disulfide bonds

None

Other bonds (neither disulfide, covalent nor hydrogen bonds, e.g. Zinc–sulphur bond)

None

Non-standard residues

None

Content subtype: bmr34466_3.str

#	Content subtype	Saveframe	Status	# of rows	Experiment type	Sequence coverage (%)
1	Assigned chemical shifts	assigned_chemical_shifts_1	Warning	2240		98.0 (chain: 1, length: 205)
1	Chemical shift references	chem_shift_reference_1	OK	3		No information

Assigned chemical shifts

Saveframe: assigned_chemical_shifts_1
- Status: Warning
- Sequence coverage of experimental data
  - Entity_assembly_ID: 1, Chain length: 205, Sequence coverage: 98.0%
- Total number of assignments
  - ¹H chemical shifts: 1370
  - ¹³C chemical shifts: 655
  - ¹⁵N chemical shifts: 215
- Completeness of assignments
  - Entity_assemble_ID: 1
- Redox state of CYS
- Peptide bond of PRO
- Tautomeric state of HIS
- Rotameric state of ILE , LEU, and VAL
- Histogram of normalized assigned chemical shifts
- Random coil index and derived parameters (S² and NMR RMSD)
  - Chain_ID: 1

Keywords 3'-splice site, SPLICING, polypyrimidine tract, splicing regulation