Zambon Alexander C, Zhang Lingzhi, Minovitsky Simon, Kanter Joan R, Prabhakar Shyam, Salomonis Nathan, Vranizan Karen, Dubchak Inna, Conklin Bruce R, Insel Paul A
Gladstone Institute of Cardiovascular Disease, University of California, San Francisco, CA 94141, USA.
Proc Natl Acad Sci U S A. 2005 Jun 14;102(24):8561-6. doi: 10.1073/pnas.0503363102. Epub 2005 Jun 6.
Although a substantial number of hormones and drugs increase cellular cAMP levels, the global impact of cAMP and its major effector mechanism, protein kinase A (PKA), on gene expression is not known. Here we show that treatment of murine wild-type S49 lymphoma cells for 24 h with 8-(4-chlorophenylthio)-cAMP (8-CPT-cAMP), a PKA-selective cAMP analog, alters the expression of approximately 4,500 of approximately 13,600 unique genes. By contrast, gene expression was unaltered in Kin- S49 cells (that lack PKA) incubated with 8-CPT-cAMP. Changes in mRNA and protein expression of several cell-cycle regulators accompanied cAMP-induced G1-phase cell-cycle arrest of wild-type S49 cells. Within 2 h, 8-CPT-cAMP altered expression of 152 genes that contain evolutionarily conserved cAMP-response elements within 5 kb of transcriptional start sites, including the circadian clock gene Per1. Thus, cAMP through its activation of PKA produces extensive transcriptional regulation in eukaryotic cells. These transcriptional networks include a primary group of cAMP-response element-containing genes and secondary networks that include the circadian clock.
尽管大量激素和药物会提高细胞内的环磷酸腺苷（cAMP）水平，但cAMP及其主要效应机制——蛋白激酶A（PKA）对基因表达的整体影响尚不清楚。在此我们表明，用PKA选择性cAMP类似物8-（4-氯苯硫基）-环磷酸腺苷（8-CPT-cAMP）处理小鼠野生型S49淋巴瘤细胞24小时，会改变约13,600个独特基因中约4,500个基因的表达。相比之下，用8-CPT-cAMP孵育的Kin-S49细胞（缺乏PKA）的基因表达未发生改变。几种细胞周期调节因子的mRNA和蛋白质表达变化伴随着cAMP诱导的野生型S49细胞G1期细胞周期停滞。在2小时内，8-CPT-cAMP改变了152个基因的表达，这些基因在转录起始位点5 kb范围内包含进化上保守的cAMP反应元件，包括昼夜节律时钟基因Per1。因此，cAMP通过激活PKA在真核细胞中产生广泛的转录调控。这些转录网络包括一组主要的含cAMP反应元件的基因和包括昼夜节律时钟在内的二级网络。
