A rapid, PPAR-γ-dependent effect of pioglitazone on the phosphorylation of MYPT

Kevin B. Atkins, Brittany Irey, Nan Xiang, Frank C. Brosius

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Peroxisome proliferator-activated receptor (PPAR)-γ ligands, thiazolidinediones, have been demonstrated to regulate vascular reactivity. We examined the effect of pioglitazone (PIO; 20 μM) in rat primary cultured aortic smooth muscle cells on constitutive phosphorylation of the regulatory subunit of myosin phosphatase (MYPT). PIO decreased the phosphorylation of Thr697 on MYPT within 15 min, and the inhibition was maintained up to 6 h. The PPAR-γ antagonist GW-9662 (5 μM) abrogated the inhibition of Thr697 phosphorylation mediated by PIO. Because longer-term PIO treatment inhibits RhoA/Rho kinase (ROCK) signaling and Thr697 phosphorylation, we tested the effect of the ROCK inhibitor Y-27632 (10 μM) on the inhibition of Thr697 phosphorylation by PIO. Y-27632 alone inhibited Thr697 phosphorylation, and there was an additive effect with PIO. In addition, up to 1 h of PIO treatment did not affect RhoA localization or decrease ROCK-dependent phosphorylation of Thr855. These results suggest that the effect of PIO is independent of inhibition of RhoA/ROCK. PIO increased the phosphorylation of Ser696 in the same time course as its effect on Thr697. Ser696 has been shown to be phosphorylated by PKA and PKG. PKA inhibitor H-89 (10 μM) and PKG inhibitor KT-5823 (0.5 μM) abrogated the effect of PIO on both Thr 697 and Ser696 phosphorylation. The constitutive turnover of phosphorylation of Thr697 is rapid, suggesting that the decreased phosphorylation of Thr697 by PIO is due to enhanced phosphorylation of Ser696. This is supported by the finding that PIO blocks ANG II-stimulated phosphorylation of Thr697 but not ANG II-stimulated RhoA translocation. Therefore, the effect of shorter-term PIO apparently is to increase myosin light chain phosphatase activity, thereby desensitizing the vascular smooth muscle to agonist signaling.

Original languageEnglish (US)
Pages (from-to)C1151-C1161
JournalAmerican Journal of Physiology - Cell Physiology
Volume296
Issue number5
DOIs
StatePublished - May 2009
Externally publishedYes

Keywords

  • Myosin phosphatase regulatory subunit
  • Peroxisome proliferator-activated receptor-γ
  • Vascular

ASJC Scopus subject areas

  • Physiology
  • Cell Biology

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