Meisner H, Loose D S, Hanson R W
Biochemistry. 1985 Jan 15;24(2):421-5. doi: 10.1021/bi00323a027.
The effect of hormones on the transcription rate of cytosolic phosphoenolpyruvate carboxykinase and level of mRNA for this enzyme in the rat kidney has been investigated. In renal nuclei isolated from rats given dibutyryladenosine cyclic 3',5'-phosphate (Bt2cAMP) or 8-bromoadenosine cyclic 3',5'-phosphate (8-Br-cAMP), [32P]UMP incorporation into hybridizable phosphoenolpyruvate carboxykinase mRNA increased severalfold within 1 h. Changes in the concentration of cytosolic phosphoenolpyruvate carboxykinase mRNA, measured by hybridization of [32P]cDNA to poly(A)+ mRNA, paralleled alterations in the transcription rate. Dexamethasone treatment of adrenalectomized rats increased the transcription rate and the level of phosphoenolpyruvate carboxykinase mRNA 3-4-fold after 4 h. Both parameters then declined to control values by 8 h. When dexamethasone (5 mg/kg) and Bt2cAMP (25 mg/kg) were given together, the rate of phosphoenolpyruvate carboxykinase RNA synthesis and the level of cytosolic mRNA were not increased more than those with either drug alone. Transcription of the gene for renal phosphoenolpyruvate carboxykinase was not affected by diabetes or glucose refeeding but was increased 2-fold after 24 h of starvation and reduced by bicarbonate feeding after 2 h. We conclude that glucocorticoids and cAMP change the rate of transcription of the phosphoenolpyruvate carboxykinase gene in rat kidney, leading to changes of similar magnitude in mRNA level and, hence, enzyme activity. The results presented here and in previous work [Lamers, W., Hanson, R. W., & Meisner, H. (1982) Proc. Natl. Acad. Sci. U.S.A. 79, 5137] indicate that the transcription rate of the gene for phosphoenolpyruvate carboxykinase in liver and kidney responds to hormones in a tissue-specific manner.
研究了激素对大鼠肾脏中胞质磷酸烯醇式丙酮酸羧激酶转录速率及该酶mRNA水平的影响。在从给予二丁酰腺苷环3',5'-磷酸(Bt2cAMP)或8-溴腺苷环3',5'-磷酸(8-Br-cAMP)的大鼠分离出的肾细胞核中,[32P]UMP掺入可杂交的磷酸烯醇式丙酮酸羧激酶mRNA在1小时内增加了数倍。通过[32P]cDNA与聚(A)+mRNA杂交测量的胞质磷酸烯醇式丙酮酸羧激酶mRNA浓度变化与转录速率的改变平行。对肾上腺切除的大鼠给予地塞米松处理4小时后,磷酸烯醇式丙酮酸羧激酶的转录速率和mRNA水平增加了3至4倍。然后这两个参数在8小时时降至对照值。当同时给予地塞米松(5mg/kg)和Bt2cAMP(25mg/kg)时,磷酸烯醇式丙酮酸羧激酶RNA合成速率和胞质mRNA水平的增加不超过单独使用任何一种药物时的增加。肾脏磷酸烯醇式丙酮酸羧激酶基因的转录不受糖尿病或葡萄糖再喂养的影响,但在饥饿24小时后增加2倍,在给予碳酸氢盐2小时后降低。我们得出结论,糖皮质激素和cAMP改变大鼠肾脏中磷酸烯醇式丙酮酸羧激酶基因的转录速率,导致mRNA水平和酶活性发生相似程度的变化。本文及先前工作[拉默斯,W.,汉森,R.W.,& 迈斯纳,H.(1982年)美国国家科学院院刊79,5137]中的结果表明,肝脏和肾脏中磷酸烯醇式丙酮酸羧激酶基因的转录速率以组织特异性方式对激素作出反应。