ZFIN Image: Iwase et al., 2007, Fig. 5

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Figure Caption

Fig. 5 Recognition of H3K9me3 by a PHD Finger of SMCX

(A) Histone-peptide binding assay. Two PHD fingers of SMCX, referred as PHD1 and PHD2, were fused to GST and purified from bacteria. Biotinylated histone peptides shown in a panel were incubated with purified PHD fingers, and pulled down by streptavidin agarose. After washing, the bound GST fusion proteins were visualized by western blot analysis using anti-GST antibody. Among the differently methylated histone peptides that we tested, only H3K9me3 peptide bound PHD1. No detectable binding was found for PHD2.

(B) A388P mutation associated with X-linked mental retardation affects PHD1 binding to H3K9me3. GST-fused wild-type and mutant PHD1 carrying A388P were incubated with H3K9me3 peptide and washed four times, and the bound fractions were analyzed by western blot analysis.

Acknowledgments

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Reprinted from Cell, 128(6), Iwase, S., Lan, F., Bayliss, P., de la Torre-Ubieta, L., Huarte, M., Qi, H.H., Whetstine, J.R., Bonni, A., Roberts, T.M., and Shi, Y., The X-linked mental retardation gene SMCX/JARID1C defines a family of histone H3 lysine 4 demethylases, 1077-1088, Copyright (2007) with permission from Elsevier. Full text @ Cell