4WSA

Crystal structure of Influenza B polymerase bound to the vRNA promoter (FluB1 form)

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	2014-04-04
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9730
Spacegroup name	P 32 2 1
Unit cell lengths	199.700, 199.700, 252.680
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	48.978 - 3.400
R-factor	0.2304
Rwork	0.229
R-free	0.26530
Structure solution method	SIRAS
RMSD bond length	0.003
RMSD bond angle	0.657
Data reduction software	XDS
Data scaling software	XSCALE
Phasing software	PHASER
Refinement software	PHENIX ((phenix.refine: dev_1760))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	3.520
High resolution limit [Å]	3.400	3.400
Rmerge	0.121	1.300
Number of reflections	79266
<I/σ(I)>	12.6	1.5
Completeness [%]	98.5	89.9
Redundancy	11.8	5.8

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	9	281	FluB polymerase was concentrated to 9 mg per ml (37 microM) in a buffer containing 500 mM NaCl, 50 mM Hepes pH 7.5, 5% glycerol and 2mM Tris(2-carboxyethyl)phosphine (TCEP), and mixed with 40 microM vRNA for crystallization in hanging drops at 4 C. A trigonal crystal form (FluB1) was obtained by mixing polymerase with nucleotides 5-18 of the 3 prime end and 1-14 of the 5 prime end of the vRNA in a condition containing 0.1 M bicine pH 9.0, 10% MPD.