Heat shock enhances CMV-IE promoter-driven metabotropic glutamate receptor expression and toxicity in transfected cells
Heat shock enhances CMV-IE promoter-driven metabotropic glutamate receptor expression and toxicity in transfected cells
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DOI：
10.1016/j.neuropharm.2011.01.010
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发表时间：
2011-06-01
影响因子：
4.7
通讯作者：
Wroblewski, Jarda T.
中科院分区：
文献类型：
--
作者：
Pshenichkin, Sergey;Surin, Alexander;Wroblewski, Jarda T.
在CHO-K1细胞中，热休克强烈激活由巨细胞病毒即刻早期(CMV-IE)启动子驱动的报告基因的表达。热休克处理(42.5℃，2小时)显著提高了核提取液中启动子的DNA结合活性。在CMV启动子控制下表达mGluR1a和mGluR5a受体的CHO细胞中，热休克使受体蛋白表达、mRNA水平和受体功能增加，通过PI水解度、细胞内Ca(2+)和cAMP来评价。高温使钙离子反应的平均幅度、反应细胞数增加，并揭示了mGluR1a受体的毒性特性。热休克还可有效增加EGFR的表达，因此，热休克对中国仓鼠卵巢细胞mGluR表达和功能的影响可能归因于CMV启动子的激活。此外，这种影响并不局限于CHO细胞，因为热休克也增加了PC-12和HEK293细胞中EGFP的表达。热休克处理可能是研究在CMV启动子控制下的异源系统中表达的蛋白质的功能的有用工具。这对于增加瞬时转基因中的蛋白质表达，在药物筛选应用中增强受体的表达，以及控制具有毒性性质的蛋白质的表达，可能特别有价值。本文是题为《神经药理学的趋势：纪念埃米尼奥·科斯塔》的特刊的一部分。(C)2011爱思唯尔有限公司。保留所有权利。
In CHO-K1 cells, heat shock strongly activated reporter-gene expression driven by the cytomegalovirus immediate-early (CMV-IE) promoter from adenoviral and plasmid vectors. Heat shock treatment (2 h at 42.5 degrees C) significantly enhanced the promoter DNA-binding activity in nuclear extracts. In CHO cells expressing mGluR1a and mGluR5a receptors under the control of the CMV promoter, heat shock increased receptor protein expression, mRNA levels and receptor function estimated by measurement of PI hydrolysis, intracellular Ca(2+) and cAMP. Hyperthermia increased average amplitudes of Ca(2+) responses, the number of responding cells, and revealed the toxic properties of mGluR1a receptor. Heat shock also effectively increased the expression of EGFR Hence, heat shock effects on mGluR expression and function in CHO cells may be attributed to the activation of the CMV promoter. moreover, this effect was not limited to CHO cells as heat shock also increased EGFP expression in PC-12 and HEK293 cells. Heat shock treatment may be a useful tool to study the function of proteins expressed in heterologous systems under control of the CMV promoter. It may be especially valuable for increasing protein expression in transient transfections, for enhancing receptor expression in drug screening applications and to control the expression of proteins endowed with toxic properties.This article is part of a Special Issue entitled 'Trends in Neuropharmacology: In Memory of Erminio Costa'. (C) 2011 Elsevier Ltd. All rights reserved.