Soluble adenylyl cyclase links Ca2+ entry to Ca2+/cAMP-response element binding protein (CREB) activation in vascular smooth muscle

Sci Rep. 2019 May 13;9(1):7317. doi: 10.1038/s41598-019-43821-3.

Abstract

Ca2+-transcription coupling controls gene expression patterns that define vascular smooth muscle cell (VSMC) phenotype. Although not well understood this allows normally contractile VSMCs to become proliferative following vessel injury, a process essential for repair but which also contributes to vascular remodelling, atherogenesis and restenosis. Here we show that the Ca2+/HCO3--sensitive enzyme, soluble adenylyl cyclase (sAC), links Ca2+ influx in human coronary artery smooth muscle cells (hCASMCs) to 3',5'-cyclic adenosine monophosphate (cAMP) generation and phosphorylation of the transcription factor Ca2+/cAMP response element binding protein (CREB). Store-operated Ca2+ entry (SOCE) into hCASMCs expressing the FRET-based cAMP biosensor H187 induced a rise in cAMP that mirrored cytosolic [Ca2+]. SOCE also activated the cAMP effector, protein kinase A (PKA), as determined by the PKA reporter, AKAR4-NES, and induced phosphorylation of vasodilator-stimulated phosphoprotein (VASP) and CREB. Transmembrane adenylyl cyclase inhibition had no effect on the SOCE-induced rise in cAMP, while sAC inhibition abolished SOCE-generated cAMP and significantly reduced SOCE-induced VASP and CREB phosphorylation. This suggests that SOCE in hCASMCs activates sAC which in turn activates the cAMP/PKA/CREB axis. sAC, which is insensitive to G-protein modulation but responsive to Ca2+, pH and ATP, may therefore act as an overlooked regulatory node in vascular Ca2+-transcription coupling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Methyl-3-isobutylxanthine / pharmacology
  • Adenylyl Cyclases / metabolism*
  • Calcium / metabolism*
  • Calcium Signaling / drug effects
  • Cations, Divalent / metabolism
  • Cell Line
  • Colforsin / pharmacology
  • Coronary Vessels / cytology
  • Cyclic AMP / metabolism
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Estradiol / analogs & derivatives
  • Estradiol / pharmacology
  • Humans
  • Muscle, Smooth, Vascular / cytology
  • Muscle, Smooth, Vascular / metabolism*
  • Myocytes, Smooth Muscle / metabolism*
  • Phosphorylation / drug effects
  • Thapsigargin / pharmacology
  • Transcriptional Activation / drug effects

Substances

  • CREB1 protein, human
  • Cations, Divalent
  • Cyclic AMP Response Element-Binding Protein
  • Colforsin
  • Estradiol
  • Thapsigargin
  • 2-hydroxyestradiol
  • Cyclic AMP
  • ADCY10 protein, human
  • Adenylyl Cyclases
  • Calcium
  • 1-Methyl-3-isobutylxanthine