6EVK

Crystal structure of bat influenza A/H17N10 polymerase with viral RNA promoter and cap analogue m7GTP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID23-1
Synchrotron site	ESRF
Beamline	ID23-1
Temperature [K]	100
Detector technology	PIXEL
Collection date	2014-09-05
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.979
Spacegroup name	C 1 2 1
Unit cell lengths	269.340, 148.700, 88.510
Unit cell angles	90.00, 98.17, 90.00

Refinement procedure

Resolution	50.000 - 2.900
R-factor	0.23592
Rwork	0.234
R-free	0.27294
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4wsb
RMSD bond length	0.008
RMSD bond angle	1.139
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	REFMAC (5.8.0158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.980
High resolution limit [Å]	2.900	2.900
Rmeas	0.115	1.380
Number of reflections	76144
<I/σ(I)>	10.75	1.08
Completeness [%]	99.6	99.8
Redundancy	4.63	4.65
CC(1/2)	0.998	0.597

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	5	281	Bat influenza polymerase protein in 50 mM HEPES-NaOH, 500 mM NaCl, 5 % glycerol, 2 mM TCEP, pH = 7.5 was adjusted to a concentration of 10 mg per ml, mixed in a 1:1 ratio with vRNA, which was an equimolar mixture of nucleotides 1-16 from the 5 prime end and nucleotides 1-18 or 3-18 from the 3 prime end. Protein-RNA with the addition of 5 mM m7GTP was mixed with mother liquor containing 0.7-1.5 M sodium-potassium phosphate at pH 5.0