Figure 3. The ZMET2 CD recognizes H3K9me in the binding step and the BAH domain recognizes H3K9me in the catalytic step.
(A) WT, F441A and W224L ZMET2 affinity for fluorescein-labeled H3 tail peptides measured by fluorescence polarization. (For these experiments, n=2).
(B) WT, F441A and W224L ZMET2 affinity for WT and H3Kc9me3 mononucleosomes measured by fluorescence polarization. Experiments with WT and F441A (CDx) ZMET2 were performed in duplicate (n=2). Experiments with W224L (BAHx) were performed in quadruplicate (n=4).
(C) DNA methyltransferase activity for WT, F441A and W224L ZMET2 on WT and H3Kc9me3 dinucleosomes. Reactions were conducted under single-turnover conditions in which the concentration of each ZMET2 protein was in excess and saturating over the dinucleosome concentration. (For these experiments, n=2).