7Z42

Influenza B polymerase with Pol II pSer5 CTD peptide mimic bound in site 2B

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	2017-09-25
Detector	DECTRIS PILATUS 6M-F
Wavelength(s)	1.25422
Spacegroup name	P 1 21 1
Unit cell lengths	130.930, 202.269, 135.733
Unit cell angles	90.00, 110.54, 90.00

Refinement procedure

Resolution	127.106 - 2.418
Rwork	0.223
R-free	0.25670
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5fmz
RMSD bond length	0.003
RMSD bond angle	1.197
Data reduction software	autoPROC
Data scaling software	autoPROC
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	127.106	2.690
High resolution limit [Å]	2.418	2.420
Rmerge	0.112	0.618
Rmeas	0.136	0.769
Number of reflections	154519	7573
<I/σ(I)>	7.7	1.9
Completeness [%]	61.4	64
Redundancy	3	2.5
CC(1/2)	0.992	0.630

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	7.5	293	FluPolB PA mutant K135A at 9 mg per ml was mixed with 40 microM of nucleotides 1-13 vRNA 5prime end, 5-pAGUAGUAACAAGA-3, and 1.8 mM 28-mer CTD peptide, YSPTpSPS x 4 in a buffer containing 50 mM HEPES pH 7.5, 500 mM NaCl, 5% glycerol, 2 mM TCEP. Hanging drops for crystallization were set up at 20 C.