Liu H, Palmer D, Jimmo S L, Tilley D G, Dunkerley H A, Pang S C, Maurice D H
Departments of Pharmacology & Toxicology, Anatomy & Cell Biology, and Pathology, Queen's University, Kingston, Ontario K7L 3N6, Canada.
J Biol Chem. 2000 Aug 25;275(34):26615-24. doi: 10.1074/jbc.M001634200.
Multiple families of cyclic nucleotide phosphodiesterases (PDE) have been described, and the regulated expression of these genes in cells is complex. Although cAMP is known to control the expression of certain PDE in cells, presumably reflecting a system of feedback on cAMP signaling, relatively little is known about the influence of non-cAMP signaling systems on PDE expression. In this study, we describe a novel mechanism by which activators of the protein kinase C (PKC)-Raf-MEK-ERK cascade regulate phosphodiesterase 4D (PDE4D) expression in vascular smooth muscle cells (VSMC) and assess the functional consequences of this effect. Whereas a prolonged elevation of cAMP in VSMC resulted in a protein kinase A (PKA)-dependent induction of expression of two PDE4D variants (PDE4D1 and PDE4D2), simultaneous activation of both the cAMP-PKA and PKC-Raf-MEK-ERK signaling cascades blunted this cAMP-mediated increase in PDE4D expression. By using biochemical, molecular biological, and pharmacological approaches, we demonstrate that this PDE4D-selective effect of activators of the PKC-Raf-MEK-ERK cascade was mediated through a mechanism involving altered PDE4D mRNA stability and markedly attenuated the cAMP-mediated desensitization that results from prolonged activation of the cAMP signaling system in cells. The data are presented in the context of activators of the PKC-Raf-MEK-ERK cascade having both short and long term effects on PDE4D activity and expression in cells that may influence cAMP signaling.
已描述了多个环核苷酸磷酸二酯酶(PDE)家族,这些基因在细胞中的表达调控很复杂。虽然已知cAMP可控制细胞中某些PDE的表达,推测这反映了对cAMP信号的一种反馈系统,但关于非cAMP信号系统对PDE表达的影响了解相对较少。在本研究中,我们描述了一种新机制,蛋白激酶C(PKC)-Raf-MEK-ERK级联反应的激活剂通过该机制调节血管平滑肌细胞(VSMC)中磷酸二酯酶4D(PDE4D)的表达,并评估这种效应的功能后果。VSMC中cAMP的长时间升高导致蛋白激酶A(PKA)依赖性诱导两种PDE4D变体(PDE4D1和PDE4D2)的表达,而cAMP-PKA和PKC-Raf-MEK-ERK信号级联反应的同时激活减弱了这种cAMP介导的PDE4D表达增加。通过生化、分子生物学和药理学方法,我们证明PKC-Raf-MEK-ERK级联反应激活剂对PDE4D的这种选择性作用是通过一种涉及改变PDE4D mRNA稳定性的机制介导的,并且显著减弱了细胞中cAMP信号系统长期激活导致的cAMP介导的脱敏作用。这些数据是在PKC-Raf-MEK-ERK级联反应激活剂对细胞中PDE4D活性和表达具有短期和长期影响的背景下呈现的,这些影响可能会影响cAMP信号。