Myc-mediated SDHA Acetylation Triggers Epigenetic Regulation of Gene Expression and Tumorigenesis.

Trimethylated H3K4 (H3K4me3), which is often found surrounding the transcriptional start site of active genes, is strongly enriched in the promoter regions of Myc-targeted genes. Succinate, which inhibits JmjC domain-containing histone demethylases (JHDMs), can induce H3K4me3 level of the promoter regions. It's interesting to ask whether Myc can mediate H3K4me3 by regulating cellular succinate level. Here, We used ChIP-seq to study Myc modulates H3K4me3 by altering succinate levels, dimethyl succinate (DMS), a membrane-permeable succinate analog, was introduced to treat CA46 cells with high or depleted Myc expression. And we also performed RNA-seq assays in Myc expressed or depleted CA46 cells with or without DMS treatment. Overall design: CA46 cells were infected with viruses expressing non-targeting control (NTC) or Myc shRNA for 48h in presence of methyl succinate (DMS) and followed by H3K4me3 ChIP-seq to address whether succinate can induce H3K4me3 level of the promoter regions in CA46 cells with depleted Myc expression. For the CA46 cells' ChIP-seq experiments with spike-in control, Drosophila chromatin (Active Motif 53083) and a Drosophila-specific antibody (Active Motif 61686) were used for each reaction. The D. melanogaster-specific antibody is used to immunoprecipitate D. melanogaster chromatin from each reaction, which is then used as a reference for normalization during data analysis. And ChIP-seq was sequenced at Novogene (Tianjin, China) on the Illumina novaseq6000 platform. Meanwhile, we also performed RNA extraction from these cell samples and sequenced on Illumina novaseq6000 platform.