4TQS

Ternary complex of Y-family DNA polymerase Dpo4 with (5'S)-8,5'-Cyclo-2'-deoxyguanosine and dCTP

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	APS BEAMLINE 21-ID-F
Synchrotron site	APS
Beamline	21-ID-F
Temperature [K]	110
Detector technology	CCD
Collection date	2014-03-28
Detector	MARMOSAIC 225 mm CCD
Wavelength(s)	1.0
Spacegroup name	P 1 21 1
Unit cell lengths	52.120, 184.242, 52.124
Unit cell angles	90.00, 110.09, 90.00

Refinement procedure

Resolution	28.407 - 2.060
R-factor	0.2312
Rwork	0.228
R-free	0.28990
RMSD bond length	0.009
RMSD bond angle	1.147
Data scaling software	HKL-2000
Phasing software	MOLREP
Refinement software	PHENIX ((phenix.refine: 1.9_1690))

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	28.407	2.100
High resolution limit [Å]	2.060	2.060
Number of reflections	58860
<I/σ(I)>		3
Completeness [%]	98.5	98.5
Redundancy		3.4

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP		291	220 microM Dpo4, 264 microM primer-template DNA complex, 5 mM MgCl2, 1 mM dTTP, 20 mM Tris-HCl (pH 7.4), 60 mM NaCl, 2% glycerol (v/v), and 5 mM 2-mercaptoethanol were incubated at 37 C for 5 min. Droplets (a 1:1 mixture of Dpo4-DNA complex mixture and the reservoir solution, v/v) were equilibrated against a reservoir solution containing 0.1M Tris-HCl (pH 7.4), 15% polyethylene glycol 3350 (w/v), 0.1 M Ca(CH3COO)2, and 2% glycerol (v/v).