Mitogen-induced transcriptional programming in human fibroblasts

• Published in: Gene 2021-10, Vol.800, p.145842-145842, Article 145842
• Main Authors: Sharma, Kiran L., Jia, Shuo, Beacon, Tasnim H., Adewumi, Ifeoluwa, López, Camila, Hu, Pingzhao, Xu, Wayne, Davie, James R.
• Format: Article
• Language: English
• Subjects: Immediate-early genes; Mitogen- and stress-activated protein kinase; Protein kinase A; RNA sequencing; Transcriptome
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2021.145842

•Transcriptome analyses of human fibroblasts stimulated with EGF or TPA ± H89.•Identification of immediate-early genes induced by EGF or TPA.•Identification of immediate-early genes dependent upon the activity of MSK or PKA.•Identification of transcriptional repressor genes upregulated with H89. Treatment of serum-starved quiescent human cells with fetal bovine serum (FBS), epidermal growth factor (EGF), or the phorbol ester (12-O-tetradecanoylphorbol-13-acetate, TPA) activates the RAS-MAPK pathway which initiates a transcriptional program which drives cells toward proliferation. 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. The induction of many immediate-early genes was independent of MSK activity. However, the induction of immediate-early genes attenuated with H89 also had reduced induction with the PKA inhibitor, Rp-cAMPS. Several EGF-induced genes, coding for transcriptional repressors, were further upregulated with H89 but not with Rp-cAMPS, suggesting a role for MSK in modulating the induction level of these genes.