Figure 4
From: Alternative catalytic residues in the active site of Esco acetyltransferases
S812, D813, S773, and E728 function cooperatively in the catalysis of MmEsco1. (A) Flow cytometry profiles of G1-phase arrested MEFsEsco1+/+, MEFsEsco1−/− and MEFsEsco1−/− expressing ectopic wild-type MmEsco1. Unsynchronized MEFsEsco1−/− were used as reference. The numbers show the percentage of cells in G1, S, G2/M phase. (B) Immunoblot of MEFsEsco1+/+, MEFsEsco1−/− and MEFsEsco1−/− expressing ectopic wild-type MmEsco1, arrested in G1-phase. Of note, transiently transfected MmEsco1 was expressed at a level of about 4-fold that of endogenous MmEsco1. (C) Immunoblots of MEFsEsco1−/− transiently expressing wild-type or mutant versions of MmEsco1, arrested in G1-phase (Supplementary Fig. S4A). Note that MEFsEsco1−/− expressed comparable levels of different MmEsco1 variants. (D) Quantification of the data shown in C. The dotted line indicates the Smc3 acetylation value for MEFsEsco1−/−. Data were normalized to wild-type signal and are shown as mean ± SEM (n = 3). In all immunoblotting experiments, the chromatin-bound fractions were analyzed using a 2-fold serial dilution. Figure adapted from50.
