Elevated cyclic-AMP represses expression of exchange protein activated by cAMP (EPAC1) by inhibiting YAP-TEAD activity and HDAC-mediated histone deacetylation

• Published in: Biochimica et biophysica acta. Molecular cell research 2019-10, Vol.1866 (10), p.1634-1649
• Main Authors: Ebrahimighaei, Reza, McNeill, Madeleine C., Smith, Sarah A., Wray, Jason P., Ford, Kerrie L., Newby, Andrew C., Bond, Mark
• Format: Article
• Language: English
• Subjects: ACTIN; Actin Cytoskeleton - metabolism; Actins - metabolism; Amides; Animals; Bridged Bicyclo Compounds, Heterocyclic - metabolism; cAMP; Cell Culture Techniques; Cell Line; Cyclic AMP - metabolism; Cytochalasin D - metabolism; EPAC; Fibroblasts - metabolism; Guanine Nucleotide Exchange Factors - genetics; Guanine Nucleotide Exchange Factors - metabolism; HDAC; Histones - metabolism; Humans; Male; Protein Processing, Post-Translational; Pyridines; Rats; Rats, Sprague-Dawley; RNA, Messenger - metabolism; TEAD; Thiazolidines - metabolism; YAP
• ISSN: 0167-4889; 1879-2596
• DOI: 10.1016/j.bbamcr.2019.06.013

Ligand-induced activation of Exchange Protein Activated by cAMP-1 (EPAC1) is implicated in numerous physiological and pathological processes, including cardiac fibrosis where changes in EPAC1 expression have been detected. However, little is known about how EPAC1 expression is regulated. Therefore, we investigated regulation of EPAC1 expression by cAMP in cardiac fibroblasts. Elevation of cAMP using forskolin, cAMP-analogues or adenosine A2B-receptor activation significantly reduced EPAC1 mRNA and protein levels and inhibited formation of F-actin stress fibres. Inhibition of actin polymerisation with cytochalasin-D, latrunculin-B or the ROCK inhibitor, Y-27632, mimicked effects of cAMP on EPAC1 mRNA and protein levels. Elevated cAMP also inhibited activity of an EPAC1 promoter-reporter gene, which contained a consensus binding element for TEAD, which is a target for inhibition by cAMP. Inhibition of TEAD activity using siRNA-silencing of its co-factors YAP and TAZ, expression of dominant-negative TEAD or treatment with YAP-TEAD inhibitors, significantly inhibited EPAC1 expression. However, whereas expression of constitutively-active YAP completely reversed forskolin inhibition of EPAC1-promoter activity it did not rescue EPAC1 mRNA levels. Chromatin-immunoprecipitation detected a significant reduction in histone3-lysine27-acetylation at the EPAC1 proximal promoter in response to forskolin stimulation. HDAC1/3 inhibition partially reversed forskolin inhibition of EPAC1 expression, which was completely rescued by simultaneously expressing constitutively active YAP. Taken together, these data demonstrate that cAMP downregulates EPAC1 gene expression via disrupting the actin cytoskeleton, which inhibits YAP/TAZ-TEAD activity in concert with HDAC-mediated histone deacetylation at the EPAC1 proximal promoter. This represents a novel negative feedback mechanism controlling EPAC1 levels in response to cAMP elevation. •Elevated cAMP reduces expression of EPAC1 in cardiac fibroblasts.•EPAC1 downregulation attenuates subsequent cAMP responses.•Elevated cAMP reduces expression of EPAC1 via two mechanisms.•Inhibition of actin-polymerisation and YAP/TAZ-TEAD activity.•HDAC1/3-mediated histone-3 lysine-27-deactelyation of the EPAC1 promoter region.