Yeast CAF-1 Assembles Histone (H3-H4) 2 Tetramers Prior to DNA Deposition

While conducting a follow-up study further detailing the mechanism of CAF-1 mediated histone deposition, we were unable to reproduce the stoichiometry measurements in Fig. 4b-d, and Fig. 5 in Winkler et al. These data originally led us to conclude that one CAF-1 molecule binds one (H3-H4)2 tetramer. We now realize that traditional stoichiometry plots are not ideal, as the determination of the inflection point relies quite heavily on a very small portion of the data. Using a number of complementary approaches (i.e. Job Plots, AUC, and SEC-MALS), we are now certain that one CAF-1 complex binds only ONE H3-H4. The correct stoichiometries have now been recently published by ourselves and one other group (1,2).