6QCV

Crystal structure of influenza B polymerase initiation state with capped 14-mer RNA primer and CTP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ESRF BEAMLINE ID29
Synchrotron site	ESRF
Beamline	ID29
Temperature [K]	100
Detector technology	PIXEL
Collection date	2018-08-24
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	0.977
Spacegroup name	P 32 2 1
Unit cell lengths	200.191, 200.191, 256.208
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	50.000 - 3.240
R-factor	0.20995
Rwork	0.209
R-free	0.23448
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5msg
RMSD bond length	0.002
RMSD bond angle	1.133
Data reduction software	XDS
Data scaling software	autoPROC
Phasing software	PHASER
Refinement software	REFMAC (5.8.0238)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	3.480
High resolution limit [Å]	3.240	3.240
Rmerge	0.175	1.190
Rmeas	0.189	1.280
Number of reflections	64019
<I/σ(I)>	7.6	1.5
Completeness [%]	67.9	60.8
Redundancy	6.9	7.3
CC(1/2)	0.996	0.650

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	4.4	277	Influenza B polymerase at 9 mg per ml in was mixed with 40 microM vRNA 5 prime end 14-mer, 40 microM vRNA 3 prime end 21-mer and 80 microM 14-mer capped RNA. The mother liquor contained 200 mM di-ammonium phosphate and 100 mM sodium acetate between pH 4.0 and 4.4. CTP was soaked for 18 hours into grown crystals at a final concentration of 5 mM in the drop.