Comparison of ATPase motor F DNA binding abilities of wild-type and variant Belinostat PXD101 proteins A Chd1142 939 16 FAM-labeled DNA Doppelstr length BP with native PAGE. Protein concentrations were 1.7, 7, 28, 110 M, and the labeled DNA was 25 nM. See also Figure S4. Hauk et al. Mol Cell page 21 Author manuscript, increases available in PMC 10th September 2011. PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH Figure 6 The interface Chromodom Ne ATPase affect both positively and negatively nucleosome sliding activity Tstests mononucleosome sliding CHD1 proteins With wild-type and mutant Chromodom Ne ATPase boundary Chen Residues Walls. From the end positioned mononucleosomes, centering this test reports of nucleosomes as a shift to nucleosome bands.
End positioned mononucleosomes were incubated with N and CHD1 CHD1 chromosomal GSK256066 proteins For 60 minutes or 939 Chd1142 proteins For 180 minutes and gel St by native PAGE. The protein concentrations of N and CHD1 CHD1 chromosomal proteins Were 0.1, 1, 10 and 100 nM, and 939 Chd1142 concentrations were 0.01, 0.1, 1 and 10 data are repr Sentative Mr. erf Leads or three times more often. The interruption of the interface Chromodom Ne ATPase partially relieves the requirement histone H4 tail. Both wild-type and H4 tail recombinant S. cerevisiae histone octamers assembled 0601 60 differently labeled DNA fragments were mixed and incubated with 5 nM CHD1 proteins Indicated for the times. Shows the quantification of nucleosome sliding in and represented, respectively.
The percentage was offset as the loss of intensity t of the lower band of all B calculated Santander nucleosomes in the alley. Data are repr Performed sentative of experiments four times or more. CHD1 chromosome is the H4 to the absence of the tail. CHD1 chromo protein was incubated with wild-type nucleosomes and H4 tail above. The quantification of the nucleosome sliding was carried out and above. The 40 minute time points show averages of two measurements, and all other time points show means and standard deviations of three measurements. Hauk et al. Mol Cell page 22 Author manuscript, increases available in PMC 10th September 2011. PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH Figure 7 A model for the regulation of chromatin remodeling by CHD1 chromodomains In this model, k Can take the chromodomains a position that inhibition of the activation of the motor prevents ATPase.
The interaction with a nucleosome relieves this inhibition by stabilizing chromodomains in an unsynchronized state, the engine in a closed conformation of ATPase hydrolysis, k can achieve professional. After hydrolysis of ATP by the motor f Promotes sliding of nucleosomes. Hauk et al. Mol Cell page 23 Author manuscript, increases available in PMC 10th September 2011. PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH Hauk et al. Page 24 Table IX-ray data collection and refinement statistics of the space group collection P6122 from 94.3 , c450.1 60.5% solvent L 1 mol / 0.9792 0.9611 USS wave lengths Aufl solution scale range1 � 50.
0 3.7 50.0 3.1 99.9 72.5 completeness � RESISTANCE Redundancy 5.5 5.4 I / σ 20.9 19.0 6.9 8.0 Refinement Statistics resolution and high varies Rsym 50.0 � 7.3 50.0 1.3 12 949 reflections 15 938 Reflections in test set R 688 841 26.14 26.66 31.83 33.56 R Free3 work2 number of atoms 5745 rms deviations Bindungsl Lengths 0.015 1.648 bond angles Ramachandran most favored statistics4 also allowed 75, 2% 22.3% 2.5% 0.0% rejected big quickly allowed 1 statistics at two resolution and high enough for the wave length tops are made of the same anisotropic diffracting crystal 2 R work given Σ | FR | Σ R 3 R has been working as a free R calculated using 5% of the data is not included in the refinement with PROCHECK 4 Calculated Mol. Author manuscript, increases available in PMC 10th September 2011. PA Author Manuscript NIH-PA Author Manuscript NIH-PA Author Manuscript NIH