HSFA1a controls plant heat stress response through its action on the 3D chromatin reorganization of enhancer-promoter interactions [ChIP-seq]

To investigate the HSFA1a, a master regulator of heat stress response, plays a key role in this reorganization by promoting the formation of promoter/enhancer chromatin loops. To determine if a typical chromatin signature could differentiate distal and proximal REs, we performed ChIP-seq experiments and analyzed the levels of four histone covalent modifications that are permissive for transcription: H3K4me3, H3K9ac, H3K18ac and H3K27ac at 0, 1 and 6 hours after heat stress on both proximal and distal REs specifically active at 1 hour after HS. In addition, To assess if RNAPII could also by recruited at distal active enhancers in plants, we performed an RNAPII ChIP-seq on plant subjected to 0, 1 and 6 hours of heat stress and analyzed its accumulation on distal REs specifically active at 1 hour after HS. Moreover, we did ChIP-seq of H3K27me1 associated constitutive heterochromatin at normal condition. Comparative analysis of ChIP-seq data for tomato M82 wild type at heat stress 0h, 1h and 6h.