5MSG

Influenza B polymerase bound to vRNA promoter and capped RNA primer

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	2015-10-02
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.2724
Spacegroup name	P 32 2 1
Unit cell lengths	200.410, 200.410, 254.610
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	173.560 - 3.800
R-factor	0.23631
Rwork	0.235
R-free	0.26745
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4wrt
RMSD bond length	0.007
RMSD bond angle	0.996
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]	173.560	4.000
High resolution limit [Å]	3.800	3.800
Rmerge	0.411	2.743
Number of reflections	58685
<I/σ(I)>	9.76	1.2
Completeness [%]	99.9	100
Redundancy	46.4	4.1
CC(1/2)	0.999	0.720

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4	277	35 microM of FluB polymerase was mixed with 40 microM of the vRNA promoter and 40 microM 13-mer capped RNA primer in a buffer containing 500 mM NaCl, 50 mM HEPES pH 7.5, 5% glycerol and 2 mM TCEP. The best diffracting crystals appeared in 100 mM sodium acetate pH 3.8 - 4.0 and 150 mM di-ammonium phosphate