1ONV

NMR Structure of a Complex Containing the TFIIF Subunit RAP74 and the RNAP II CTD Phosphatase FCP1

SOLUTION NMR

NMR Experiment
Experiment	Type	Sample Contents	Solvent	Ionic Strength	pH	Pressure	Temperature (K)	Spectrometer
1	2D NOESY	1mM cterRAP74/cterFCP1, 20mM sodium phosphate, and 1mM EDTA	100% D2O	20mM sodium phosphate buffer	6.5	ambient	300
2	3D_15N-separated_NOESY	1mM U-15N cterRAP74/cterFCP1, 20mM sodium phosphate, and 1mM EDTA	90% H2O/10% D2O	20mM sodium phosphate buffer	6.5	ambient	300
3	HNCO, (HB)CBCA(CO)NNH, HNCACB, H(CCO)NNH-TOCSY, C(CO)NNH-TOCSY	1mM U-15N,13C cterRAP74/cterFCP1, 20mM sodium phosphate, and 1mM EDTA	90% H2O/10% D2O	20mM sodium phosphate buffer	6.5	ambient	300
4	3D_15N-separated_NOESY	1mM cterRAP74/U-15N cterFCP1, 20mM sodium phosphate, and 1mM EDTA	90% H2O/10% D2O	20mM sodium phosphate buffer	6.5	ambient	300
5	HNCO, (HB)CBCA(CO)NNH, HNCACB, H(CCO)NNH-TOCSY, C(CO)NNH-TOCSY	1mM cterRAP74/U-15N,13C cterFCP1, 20mM sodium phosphate, and 1mM EDTA	90% H2O/10% D2O	20mM sodium phosphate buffer	6.5	ambient	300
6	3D_13C-separated_NOESY	1mM U-15N,C13 cterRAP74/cterFCP1, 20mM sodium phosphate, and 1mM EDTA;	100% D2O	20mM sodium phosphate buffer	6.5	ambient	300
7	2D 13C-filterd/edited NOESY	1mM U-15N,C13 cterRAP74/cterFCP1, 20mM sodium phosphate, and 1mM EDTA;	100% D2O	20mM sodium phosphate buffer	6.5	ambient	300
8	3D 13C-filterd/edited NOESY	1mM cterRAP74/U-15N,13C cterFCP1, 20mM sodium phosphate, and 1mM EDTA	100% D2O	20mM sodium phosphate buffer	6.5	ambient	300
9	3D 15N/13C separated NOESY	1mM cterRAP74/U-15N,13C cterFCP1, 20mM sodium phosphate, and 1mM EDTA	90% H2O/10% D2O	20mM sodium phosphate buffer	6.5	ambient	300

NMR Spectrometer Information
Spectrometer	Manufacturer	Model	Field Strength
1	Varian	INOVA	500
2	Varian	INOVA	600
3	Varian	INOVA	800

NMR Refinement
Method	Details	Software
torsion angle dynamics	The structures were determined based on a total of 1267 restraints, 1131 NOE-derived distance restraints (including 58 intermolecular NOE-derived distance restraints) and 136 dihedral angle restraints	CNS

NMR Ensemble Information
Conformer Selection Criteria	structures with acceptable covalent geometry,structures with the least restraint violations,structures with the lowest energy
Conformers Calculated Total Number	70
Conformers Submitted Total Number	20
Representative Model	1 (closest to the average,minimized average structure)

Additional NMR Experimental Information
Details	The structure was determined using triple-resonance NMR spectroscopy

Computation: NMR Software
#	Classification	Version	Software Name	Author
1	structure solution	CNS	1.0	Brunger
2	processing	NMRPipe		Bax
3	data analysis	PIPP		Garrett
4	data analysis	NMRView	5.0	Johnson
5	refinement	CNS	modified CNS with conformational database potential	Kay and Choy, Clores

RCSB PDB Core Operations are funded by the National Science Foundation (DBI-1832184), the US Department of Energy (DE-SC0019749), and the National Cancer Institute, National Institute of Allergy and Infectious Diseases, and National Institute of General Medical Sciences of the National Institutes of Health under grant R01GM133198.