4E54

Damaged DNA induced UV-damaged DNA-binding protein (UV-DDB) dimerization and its roles in chromatinized DNA repair

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 22-ID
Synchrotron site	APS
Beamline	22-ID
Temperature [K]	100
Detector technology	CCD
Collection date	2009-07-08
Detector	MARMOSAIC 300 mm CCD
Wavelength(s)	1.00
Spacegroup name	P 1 21 1
Unit cell lengths	76.736, 70.877, 191.448
Unit cell angles	90.00, 99.68, 90.00

Refinement procedure

Resolution	33.315 - 2.850
R-factor	0.2355
Rwork	0.233
R-free	0.28090
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3ei2
RMSD bond length	0.005
RMSD bond angle	1.354
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	SOLVE
Refinement software	PHENIX ((phenix.refine: 1.7.1_743))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.960
High resolution limit [Å]	2.850	2.850
Rmerge	0.120	0.120
Number of reflections	47108
<I/σ(I)>	12.28	7.7
Completeness [%]	89.7	67.6
Redundancy	6.5	3.5

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION	7.5	277	20mM Tris pH 7.5, 2mM MgCl2, 1mM EDTA, 2mM TECP, 5% Glycerol, 0.02% azide. UV-DDB-AP24 complex (molar ratio of 1:3 UV-DDB:DNA) at 2.5 mg/mL. 'AP24' refers to synthetic DNA substrate of 24-bpr with a central abasic site mimic., VAPOR DIFFUSION, temperature 277K