7U8Y

TREX1 Structural Studies Capture Small Molecule Inhibition and Implicate Novel DNA Dynamics

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU MICROMAX-007
Temperature [K]	100
Detector technology	PIXEL
Collection date	2020-02-17
Detector	DECTRIS PILATUS 200K
Wavelength(s)	1.54
Spacegroup name	H 3
Unit cell lengths	127.555, 127.555, 80.010
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	45.500 - 2.220
R-factor	0.18309
Rwork	0.181
R-free	0.22175
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	2oa8
RMSD bond length	0.009
RMSD bond angle	1.343
Data reduction software	HKL-3000
Data scaling software	HKL-3000
Phasing software	PHASER (v2.1.2)
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.290
High resolution limit [Å]	2.220	2.220
Rmerge	0.073	0.328
Rmeas	0.077	0.355
Rpim	0.025	0.133
Number of reflections	23262	1804
<I/σ(I)>	30.8	5.8
Completeness [%]	99.1	89.5
Redundancy	9.4
CC(1/2)	0.997	0.953

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP		298	Protein was dialyzed into 20 mM MES (pH 6.5) with 50 mM NaCl. Complex was formed by incubating the protein at 5 mg/mL with 2 mM dAMP and 5 mM MnCl2. 0.3 uL protein complex at 5 mg/ml TREX1 was mixed with an equal volume of reservoir solution and placed on a bridge above 50 uL of the reservoir solution. mTREX1-dAMP crystals grew in initial conditions of 0.2 M MES monohydrate, sodium hydroxide (pH 6.2), and 20% w/v polyethylene glycol 4,000. All crystals grew within one week. After crystal formation, 4 mM TIM009 was soaked into mTREX1-dAMP crystals for 5 days. Prior to data collection crystals were dipped into reservoir solution containing 15% glycerol in preparation for cryo-cooling