Mitogen induced-nucleosomal response pathway and transcriptional programming in mammalian fibroblasts

Stimulation of the RAS-MAPK pathway activates mitogen- and stress-activated kinases (MSK) 1 and 2, which phosphorylate histone H3 at S10 (H3S10ph) or S28 (H3S28ph) (nucleosomal response) located at the regulatory regions of immediate-early genes, setting in motion a series of chromatin remodeling events that result in transcription initiation. To investigate immediate-early genes regulated by the MSK, we have completed transcriptome analyses (RNA Sequencing) of human normal fibroblast cells (CCD-1070Sk) stimulated with EGF or TPA ± H89, a potent MSK/PKA inhibitor. Overall design: Treatment of quiescent human cells with fetal bovine serum (FBS), epidermal growth factor (EGF) or the phorbol ester (12-O-tetradecanoylphorbol-13-acetate, TPA) ± H89 following RNA Sequencing.