Phosphodiesterase 3A (PDE3A) deletion suppresses proliferation of cultured murine vascular smooth muscle cells (VSMCs) via inhibition of mitogen-activated protein kinase (MAPK) signaling and alterations in critical cell cycle regulatory proteins.

Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE

Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology

Gene Deletion*; Cell Cycle Proteins/*metabolism ; Cyclic Nucleotide Phosphodiesterases, Type 3/*deficiency ; Cyclic Nucleotide Phosphodiesterases, Type 3/*genetics ; MAP Kinase Signaling System/*genetics ; Mitogen-Activated Protein Kinases/*metabolism ; Muscle, Smooth, Vascular/*cytology; Animals ; Biocatalysis/drug effects ; Cell Cycle/drug effects ; Cell Cycle/genetics ; Cell Cycle Proteins/genetics ; Cell Proliferation/drug effects ; Cells, Cultured ; Cyclic AMP Response Element-Binding Protein/metabolism ; Cyclic AMP-Dependent Protein Kinases/metabolism ; DNA/biosynthesis ; Dual Specificity Phosphatase 1/metabolism ; Extracellular Signal-Regulated MAP Kinases/metabolism ; Gene Expression Regulation, Enzymologic/drug effects ; Gene Expression Regulation, Enzymologic/genetics ; Gene Knockout Techniques ; MAP Kinase Signaling System/drug effects ; Male ; Mice ; Muscle, Smooth, Vascular/drug effects ; Muscle, Smooth, Vascular/enzymology ; Muscle, Smooth, Vascular/metabolism ; Phosphatidylinositol 3-Kinases/metabolism ; Phosphorylation/drug effects ; Phosphorylation/genetics ; Platelet-Derived Growth Factor/pharmacology ; Proto-Oncogene Proteins c-akt/metabolism ; Proto-Oncogene Proteins c-raf/metabolism ; RNA, Small Interfering/genetics

Cyclic nucleotide phosphodiesterase 3 (PDE3) is an important regulator of cyclic adenosine monophosphate (cAMP) signaling within the cardiovascular system. In this study, we examined the role of PDE3A and PDE3B isoforms in regulation of growth of cultured vascular smooth muscle cells (VSMCs) and the mechanisms by which they may affect signaling pathways that mediate mitogen-induced VSMC proliferation. Serum- and PDGF-induced DNA synthesis in VSMCs grown from aortas of PDE3A-deficient (3A-KO) mice was markedly less than that in VSMCs from PDE3A wild type (3A-WT) and PDE3B-deficient (3B-KO) mice. The reduced growth response was accompanied by significantly less phosphorylation of extracellular signal-regulated kinase (ERK) in 3A-KO VSMCs, most likely due to a combination of greater site-specific inhibitory phosphorylation of Raf-1(Ser-²⁵⁹) by protein kinase A (PKA) and enhanced dephosphorylation of ERKs due to elevated mitogen-activated protein kinase phosphatase 1 (MKP-1). Furthermore, 3A-KO VSMCs, compared with 3A-WT, exhibited higher basal PKA activity and cAMP response element-binding protein (CREB) phosphorylation, higher levels of p53 and p53 phosphorylation, and elevated p21 protein together with lower levels of Cyclin-D1 and retinoblastoma (Rb) protein and Rb phosphorylation. Adenoviral overexpression of inactive CREB partially restored growth effects of serum in 3A-KO VSMCs. In contrast, exposure of 3A-WT VSMCs to VP16 CREB (active CREB) was associated with inhibition of serum-induced DNA synthesis similar to that in untreated 3A-KO VSMCs. Transfection of 3A-KO VSMCs with p53 siRNA reduced p21 and MKP-1 levels and completely restored growth without affecting amounts of Cyclin-D1 and Rb phosphorylation. We conclude that PDE3A regulates VSMC growth via two complementary pathways, i.e. PKA-catalyzed inhibitory phosphorylation of Raf-1 with resulting inhibition of MAPK signaling and PKA/CREB-mediated induction of p21, leading to G₀/G₁ cell cycle arrest, as well as by increased accumulation of p53, which induces MKP-1, p21, and WIP1, leading to inhibition of G₁ to S cell cycle progression.

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United States Intramural NIH HHS

0 (Cell Cycle Proteins)
0 (Cyclic AMP Response Element-Binding Protein)
0 (Platelet-Derived Growth Factor)
0 (RNA, Small Interfering)
9007-49-2 (DNA)
EC 2.7.1.- (Phosphatidylinositol 3-Kinases)
EC 2.7.11.1 (Proto-Oncogene Proteins c-akt)
EC 2.7.11.1 (Proto-Oncogene Proteins c-raf)
EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
EC 2.7.11.24 (Extracellular Signal-Regulated MAP Kinases)
EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
EC 3.1.3.48 (Dual Specificity Phosphatase 1)
EC 3.1.4.17 (Cyclic Nucleotide Phosphodiesterases, Type 3)

Date Created: 20110603 Date Completed: 20110916 Latest Revision: 20211020

20240829

PMC3138244

10.1074/jbc.M110.214155

21632535