7U4A

Crystal Structure of Zika virus xrRNA1 mutant

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-BM
Synchrotron site	APS
Beamline	22-BM
Temperature [K]	100
Detector technology	CCD
Collection date	2020-08-09
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1
Spacegroup name	P 1 21 1
Unit cell lengths	48.421, 39.137, 66.583
Unit cell angles	90.00, 94.34, 90.00

Refinement procedure

Resolution	33.200 - 3.150
R-factor	0.2168
Rwork	0.214
R-free	0.27040
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	5tpy
RMSD bond length	0.002
RMSD bond angle	0.526
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHENIX
Refinement software	PHENIX (1.18.2_3874)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	33.200	33.200	3.260
High resolution limit [Å]	3.150	7.460	3.170
Rmerge	0.114
Rmeas	0.136
Rpim	0.074
Number of reflections	4391	344	342
<I/σ(I)>	9.9
Completeness [%]	95.7
Redundancy	3.6
CC(1/2)	0.990	0.991	0.984

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		293	RNA was exchanged and concentrated into 10 mM HEPES-KOH, pH 7.5, 2.5 mM magnesium chloride. After concentration to 7.5 mg/mL (~330 uM), RNA was diluted to 4 mg/mL, 100 mM spermidine was added to a final concentration of 0.5 mM. 0.5 uL RNA was added to 2 uL crystallization solution (50 mM sodium cacodylate, pH 6.0, 150 mM sodium chloride, 4 mM calcium chloride, 0.6 mM spermine, 36% 1,6-hexanediol) and crystallized via hanging drop vapor diffusion over 1 mL crystallization solution. Crystals formed within 24 hours at 20 degrees C and were harvested after 36 hours.