Headley Violetta V, Tanveer Rasheeda, Greene Scott M, Zweifach Adam, Port J David
Department of Medicine, Division of Cardiology, University of Colorado Health Sciences Center, Denver, CO 80262, USA.
Mol Cell Biochem. 2004 Mar;258(1-2):109-19. doi: 10.1023/b:mcbi.0000012841.03400.42.
Genes encoding numerous proto-oncogenes and cytokines, as well as a number of G-protein coupled receptors, are regulated post-transcriptionally at the level of mRNA stability. A common feature of all of these genes is the presence of A + U-rich elements (AREs) within their 3' untranslated regions. We, and others, have demonstrated previously that mRNAs encoding beta-adrenergic receptors (beta-ARs) are destabilized by agonist stimulation of the beta-AR/Galphas/adenylylcyclase pathway. However, in addition to PK-A, beta-ARs can also activate or inhibit mitogen activated kinase (MAPK) cascades, in a cell-type dependent basis. Recent evidence points to an important role for MAPKs in regulating the turnover of cytokine mRNAs, such as TNFalpha. We hypothesized that activation of MAPK's may also regulate beta-AR mRNA stability. The studies conducted herein demonstrate that generalized stimulation of MAPKs (JNK, p38) with anisomycin resulted in marked stabilization of beta-AR mRNA. Reciprocally, selective inhibition of JNK with SP600125 significantly decreased beta-AR mRNA half-life. Similarly, inhibition of the MEK/ERK pathway with either PD98059 or U0126 decreased beta-AR mRNA stability substantially. However, inhibition of p38 MAPK with SB203580 produced destabilization of beta-AR mRNA only at higher, non pharmacologically selective concentrations. In contrast to their effects on several other ARE containing mRNAs, inhibition of tyrosine kinases by genistein or PI3K by wortmannin, had no detectable effect on beta-AR mRNA stability. In summary, these results demonstrate for the first time that modulation of MAPK pathways can bi-directionally influence beta-AR mRNA stability.
编码众多原癌基因、细胞因子以及一些G蛋白偶联受体的基因,在mRNA稳定性水平上受到转录后调控。所有这些基因的一个共同特征是在其3'非翻译区内存在富含A+U的元件(AREs)。我们和其他人之前已经证明,编码β-肾上腺素能受体(β-ARs)的mRNA会因β-AR/Gαs/腺苷酸环化酶途径的激动剂刺激而不稳定。然而,除了PK-A之外,β-ARs还可以在细胞类型依赖性的基础上激活或抑制丝裂原活化激酶(MAPK)级联反应。最近的证据表明MAPKs在调节细胞因子mRNA(如TNFα)的周转中起重要作用。我们假设MAPK的激活也可能调节β-AR mRNA的稳定性。本文进行的研究表明,茴香霉素对MAPKs(JNK、p38)的普遍刺激导致β-AR mRNA显著稳定。相反,用SP600125选择性抑制JNK会显著降低β-AR mRNA的半衰期。同样,用PD98059或U0126抑制MEK/ERK途径会大幅降低β-AR mRNA的稳定性。然而,用SB203580抑制p38 MAPK仅在较高的、非药理学选择性浓度下才会导致β-AR mRNA不稳定。与它们对其他几种含有ARE的mRNA的影响相反,染料木黄酮抑制酪氨酸激酶或渥曼青霉素抑制PI3K对β-AR mRNA稳定性没有可检测到的影响。总之,这些结果首次证明MAPK途径的调节可以双向影响β-AR mRNA的稳定性。