GSE118324
Mechanism suppressing H3K9 trimethylation in pluripotent stem cells and its demise by polyQ-expanded huntingtin mutations [ChIP-seq]
We find that HTT binds ATF7IP, a regulator of the histone H3 methyltransferase SETDB1. HTT inhibits the interaction of the ATF7IP-SETDB1 complex with other heterochromatin regulators and transcriptional repressors, maintaining low levels of H3K9 trimethylation (H3K9me3) in hESCs. Conversely, loss of HTT promotes global increased H3K9me3 levels. To test whether HTT knockdown also induces enrichment of H3K9me3 marks at specific genes, we performed chromatin immunoprecipitation (ChIP)-sequencing assays of hESCs using an antibody to H3K9me3. Overall design: We performed chromatin immunoprecipitation (ChIP)-sequencing using H3K9me3 antibody (Abcam, #8898, reported suitable for ChIP) comparing H9 hESCs expressing non-targeting (NT) shRNA with H9 hESCs expressing HTT shRNA. Two biological replicates from independent experiments were analyzed.