4WSB

Bat Influenza A polymerase with bound vRNA promoter

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	2014-07-20
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.9763
Spacegroup name	C 1 2 1
Unit cell lengths	268.190, 149.320, 88.620
Unit cell angles	90.00, 98.00, 90.00

Refinement procedure

Resolution	19.996 - 2.650
R-factor	0.2179
Rwork	0.215
R-free	0.26710
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4wsa
RMSD bond length	0.004
RMSD bond angle	0.732
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	2.720
High resolution limit [Å]	2.650	2.650
Rmerge	0.082	1.439
Number of reflections	99462
<I/σ(I)>	10.9	1.14
Completeness [%]	99.2	96
Redundancy	3.5	3.5

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 was mixed with mother liquor containing 0.7-1.5 M sodium-potassium phosphate at pH 5.0