Galphaq signaling is required for Rho-dependent transcriptional activation of the cyclooxygenase-2 promoter in fibroblasts.

Slice LW; Han SK; Simon MI

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Galphaq signaling is required for Rho-dependent transcriptional activation of the cyclooxygenase-2 promoter in fibroblasts.

机译：Galphaq Rho-dependent所需信号转录的激活cyclooxygenase-2启动子在成纤维细胞。

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Previously, we demonstrated that the gastrin releasing peptide (GRP) induces cyclooxygenase-2 (COX-2) expression through a Rho-dependent, protein kinase C (PKC)-independent signaling pathway in fibroblasts (Slice et al., 1999, J Biol Chem 274:27562-27566). However, the specific role of heterotrimeric guanine nucleotide binding regulatory proteins (G-proteins) that are coupled to the GRP receptor in Rho-dependent COX-2 expression has not been elucidated. In this report, we utilize embryonic fibroblasts from transgenic mice containing double gene knock-outs (DKO) for Galpha(q/11) and Galpha(12/13) to demonstrate that COX-2 promoter activation by GRP requires Galpha(q). Furthermore, we show that GRP-dependent COX-2 gene expression, as assessed by a COX-2 reporter luciferase assay, was induced in cells lacking Galpha(12/13) but was blocked in cells that did not express Galpha(q/11). GRP-dependent COX-2 promoter induction in Galpha(q/11) deficient cells was rescued by expression of wild type Galpha(q) but blocked by inhibition of calcium signaling in calcium-free media or in cells treated with 2-aminoethoxydiphenylborate (2-APB). Co-stimulation of transfected Galpha(q/11) deficient cells with GRP and thapsigargin (TG) induced the COX-2 promoter. Activation of endogenous Rho by expression of Onco-lbc or expression of Rho A Q63L resulted in COX-2 promoter activation in Galpha(q/11) deficient cells. Inhibition of Rho by Clostridium botulinum C3 toxin blocked COX-2 promoter induction. Expression of Galpha(q) Q209L in the well-characterized fibroblast cell line, NIH3T3, induced the COX-2 promoter which was blocked by expression of C3 toxin. These results demonstrate that calcium signaling mediated by Galpha(q) and Rho play critical roles in GRP-dependent COX-2 expression in fibroblasts.

机译：以前,我们证明了胃泌激素释放肽(GRP)诱发cyclooxygenase-2通过Rho-dependent (cox - 2)表达,蛋白激酶C (PKC)独立信号在成纤维细胞通路(片et al ., 1999, J临床生物化学274:27562 - 27566)。heterotrimeric鸟嘌呤核苷酸的角色绑定调节蛋白(g)耦合在Rho-dependent cox - 2 GRP受体表达尚未阐明。报告中,我们利用胚胎成纤维细胞含双转基因小鼠基因淘汰赛GRP表明cox - 2启动子的激活需要Galpha (q)。GRP-dependent cox - 2基因表达,如评估cox - 2记者荧光素酶测定,是诱导在细胞缺乏Galpha(12/13),但被阻塞细胞不表达Galpha (q / 11)。GRP-dependent cox - 2启动子感应Galpha (q / 11)缺乏细胞获救表达野生型Galpha (q),但被在calcium-free抑制钙信号媒体或在细胞治疗转染Galpha Co-stimulation (q / 11)有缺陷的细胞和GRP thapsigargin (TG)诱导cox - 2启动子。内源性ρOnco-lbc或的表情ρQ63L导致cox - 2的表达启动子的激活在Galpha (q / 11)不足细胞。C3 cox - 2启动子诱导毒素阻塞。表达Galpha (q) Q209L良好的纤维母细胞细胞系,NIH3T3被诱导的cox - 2启动子C3毒素的表情。钙信号由Galpha (q)ρGRP-dependent cox - 2中扮演关键的角色成纤维细胞中表达。

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《Journal of Cellular Physiology》 |2003年第2期|127-138|共12页
作者
Slice LW; Han SK; Simon MI;
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作者单位

Division of Digestive Diseases, Department of Medicine, CURE: Digestive Diseases Research Center, Greater Los Angeles VA Medical Center, University of California, Los Angeles, California 90095, USA. lslice@mednet.ucla.edu;

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正文语种英语
中图分类细胞生物学;
关键词
Isoenzymes; Promoter; Calcium SignalingCyclooxygenase 2FibroblastHeterotrimeric GTP-Binding Proteins;

机译：GTP-Binding蛋白质;

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Galphaq signaling is required for Rho-dependent transcriptional activation of the cyclooxygenase-2 promoter in fibroblasts.

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