FIGURE 3: The target of Mst2 in heterochromatin is not Brl1. (A) Scheme depicting the described Mst2C pathway [31] involving Brl1-K242 acetylation and H2B ubiquitylation and a potential alternative pathway on HC silencing; black arrows represent positive regulation and red lines represent negative regulation. (B) ChIP-qPCR analysis for H2B-K119ub in wild-type cells. Circles and horizontal lines represent individual data and median from 3 (left panel) and 4 independent experiments (right panel). Input-normalized ChIP data are shown relative to the median of the ChIP signals for act1⁺ (mid). (C, D) ChIP-qPCR analysis for H2B-K119ub in mutants affecting Brl1 acetylation. Circles and horizontal lines represent individual data and median from 3 independent experiments (except: WT, set2Δ: n = 4). ChIP analysis as in (B), except that ChIP data were corrected for variation in IP efficiency by normalizing to act1⁺ (mid). Note that H2B-K119ub at act1⁺ is not largely affected by Set2. (E) RT-qPCR analysis of transcript levels at pericentromeric and subtelomeric HC. Data analysis as in Figure 1C (n = 3 individual experiments).

31. Flury V, Georgescu PR, Iesmantavicius V, Shimada Y, Kuzdere T, Braun S, and Bühler M (2017). The Histone Acetyltransferase Mst2 Protects Active Chromatin from Epigenetic Silencing by Acetylating the Ubiquitin Ligase Brl1. Mol Cell 67(2): 294–307.e9. doi: 10.1016/j.molcel.2017.05.026