Fig. 6: SETD5–NCoR–HDAC3 complex restricts CBP recruitment to primed enhancers.

a Venn diagram (left) and heatmap (right) representation of genomic regions occupied by SETD5-V5 and p300. Approximately 64% of p300 genomic binding regions were co-occupied by SETD5-V5. Heatmap analysis showing ChIP-seq data for SETD5-V5 and p300 at a 20-kb region centered on SETD5-V5 binding region. b Genome browser representation for SETD5-V5, p300, and H3K27ac on Cebpa and Pparg genes in 3T3-L1 preadipocytes. Tracks 1-2 and 5-7 are the same data as Fig. 5b. c, d, e ChIP-qPCR analyses of CBP on Cebpa and Pparg genes during the early adipogenesis. 3T3-L1 preadipocytes transduced with control empty virus or SETD5 were subjected to ChIP-qPCR analyses using anti-CBP antibody c. 3T3-L1 preadipocytes transduced with control empty virus, wild-type SETD5, or Δ437-918 mutant were subjected to ChIP-qPCR analysis using anti-CBP antibody d. 3T3-L1 preadipocytes transfected with control siRNA or siRNA targeting to Setd5 were subjected to ChIP-qPCR analysis using anti-CBP antibody e. f Model of enhancer transition from primed to active state during adipogenesis. Heatmap shows the degree of recruitment of SETD5, NCoR2, HDAC3, and CBP to Cebpa and Pparg enhancers and H3K27 acetylation (based on Figs. 4d, 5c, d, 6c, Supplementary Fig. 6a). Darker color indicates more recruitment and acetylation. (i) SETD5 forms a complex with NCoR-HDAC3 and keeps a hypoacetylation state by restricting the recruitment of CBP to primed enhancers. (ii) SETD5 in NCoR-HDAC complex on primed enhancers is ubiquitinated and degraded by APC/C. (iii) Degradation of SETD5 from NCoR-HDAC3 co-repressor complex allows the unrestricted recruitment of CBP and H3K27 acetylation and transit enhancers from primed to active state. a, b ChIP-seq data for p300 were from the previously published paper³². c Data are mean ± SEM (0, 6, 12 h n = 3: 24 h n = 4; 48 h n = 5; independent experiments). Unpaired two-tailed Student’s t-test. d, e Representative of two independent experiments. Data are mean ± SD of three technical replicates. One-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05; **p < 0.01. Source data are provided as a Source data file.