4IMJ

Novel Modifications on C-terminal Domain of RNA Polymerase II can Fine-tune the Phosphatase Activity of Ssu72

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.3
Synchrotron site	ALS
Beamline	5.0.3
Temperature [K]	100
Detector technology	CCD
Collection date	2012-04-01
Detector	ADSC QUANTUM 315r
Wavelength(s)	0.9765
Spacegroup name	P 4
Unit cell lengths	127.992, 127.992, 105.775
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	37.770 - 2.580
R-factor	0.2123
Rwork	0.211
R-free	0.24370
Structure solution method	MOLECULAR REPLACEMENT
RMSD bond length	0.010
RMSD bond angle	1.050
Data reduction software	HKL-2000
Data scaling software	HKL-2000
Phasing software	PHASER
Refinement software	BUSTER (2.10.0)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	50.000	2.620
High resolution limit [Å]	2.580	2.580
Rmerge		0.845
Number of reflections	53955
<I/σ(I)>	24.2	2
Completeness [%]	100.0	99.8
Redundancy	7.5	7.3

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	8.5	293	Optimized Ssu72-symplekin crystals were obtained with reservoir solution consisting of 12% PEG3350 (w/v) and 100 mM HEPE pH 8.5. To obtain the tertiary complex structure of Drosophila Ssu72-symplekin-CTD(phos.Ser5), crystals of Ssu72-symplekin were soaked in a mother solution containing 2 mM 19mer CTD(phos.Ser5) peptides overnight at room temperature , VAPOR DIFFUSION, SITTING DROP, temperature 293K