PDE4D and PDE4B function in distinct subcellular compartments in mouse embryonic fibroblasts.
PDE4D and PDE4B function in distinct subcellular compartments in mouse embryonic fibroblasts.
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DOI：
10.1074/jbc.m110.203604
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发表时间：
2011-04-08
期刊：
影响因子：
--
通讯作者：
Conti M
中科院分区：
文献类型：
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作者：
Blackman BE;Horner K;Heidmann J;Wang D;Richter W;Rich TC;Conti M
通过cAMP的信号传导调节大多数细胞功能。因此，cAMP的时空控制对于特异性细胞靶标的差异调节至关重要。在这里，我们研究的后果PDE4B或PDE4D基因消融cAMP信号在亚细胞水平使用小鼠胚胎成纤维细胞。PDE4B消融对cAMP的整体或整体胞质溶胶积累没有影响，但增加了近膜池中的基础和依赖于磷酸盐的cAMP。相反，PDE4D消融增强了激动剂诱导的cAMP在大量胞质溶胶以及质膜中的积累。PDE4B和PDE4D消融显著改变了异丙肾上腺素诱导的血管舒张剂刺激磷蛋白（一种膜细胞骨架成分）磷酸化的时间进程和水平。然而，通过Toll样受体信号传导的第二种膜反应仅受PDE4B消融的影响。PDE4D而非PDE4B消融显著延长cAMP反应元件结合蛋白介导的转录。这些发现表明，PDE4D和PDE4B在小鼠胚胎成纤维细胞中具有专门的功能，PDE4B控制质膜附近的离散子域中的cAMP。
Signaling through cAMP regulates most cellular functions. The spatiotemporal control of cAMP is, therefore, crucial for differential regulation of specific cellular targets. Here we investigated the consequences of PDE4B or PDE4D gene ablation on cAMP signaling at a subcellular level using mouse embryonic fibroblasts. PDE4B ablation had no effect on the global or bulk cytosol accumulation of cAMP but increased both basal and hormone-dependent cAMP in a near-membrane pool. Conversely, PDE4D ablation enhanced agonist-induced cAMP accumulation in the bulk cytosol as well as at the plasma membrane. Both PDE4B and PDE4D ablation significantly modified the time course and the level of isoproterenol-induced phosphorylation of vasodilator-stimulated phosphoprotein, a membrane cytoskeletal component. A second membrane response through Toll-like receptor signaling, however, was only affected by PDE4B ablation. PDE4D but not PDE4B ablation significantly prolonged cAMP-response element-binding protein-mediated transcription. These findings demonstrate that PDE4D and PDE4B have specialized functions in mouse embryonic fibroblasts with PDE4B controlling cAMP in a discrete subdomain near the plasma membrane.