Belazzi T, Wagner A, Wieser R, Schanz M, Adam G, Hartig A, Ruis H
Institut für Allgemeine Biochemie, Universität Wien, Austria.
EMBO J. 1991 Mar;10(3):585-92. doi: 10.1002/j.1460-2075.1991.tb07985.x.
Transcription of the CTT1 (catalase T) gene of Saccharomyces cerevisiae is controlled by oxygen via heme, by nutrients via cAMP and by heat shock. Nitrogen limitation triggers a rapid, cycloheximide-insensitive derepression of the gene. Residual derepression in a cAMP-nonresponsive mutant with attenuated protein kinase activity (bcy1 tpk1w tpk2 tpk3) demonstrates the existence of an alternative, cAMP-independent nutrient signaling mechanism. Deletion analysis using CTT1-lacZ fusion genes revealed the contribution of multiple control elements to derepression, not all of which respond to the cAMP signal. A positive promoter element responding to negative control by cAMP was inactivated by deletion of a DNA region between base pairs -340 and -364. Upstream fragments including this element confer negative cAMP control to a LEU2-lacZ fusion gene. Northern analysis of CTT1 expression in the presence or absence of heme, in RAS2+ (high cAMP) and ras2 mutant (low cAMP) strains and in cells grown at low temperature (23 degrees C) and in heat-shocked cells (37 degrees C) shows that CTT1 is only induced to an appreciable extent when at least two of the three factors contributing to its expression (oxidative stress signaled by heme, nutrient starvation (low cAMP) and heat stress) activate the CTT1 promoter.
酿酒酵母CTT1(过氧化氢酶T)基因的转录受氧气通过血红素调控,受营养物质通过cAMP调控,还受热激调控。氮限制会引发该基因快速的、对放线菌酮不敏感的去阻遏。在蛋白激酶活性减弱的cAMP无反应突变体(bcy1 tpk1w tpk2 tpk3)中仍存在残余去阻遏,这表明存在一种独立于cAMP的营养信号传导机制。使用CTT1 - lacZ融合基因进行的缺失分析揭示了多个调控元件对去阻遏的作用,并非所有这些元件都对cAMP信号作出反应。通过缺失碱基对 - 340和 - 364之间的DNA区域,可使一个对cAMP负调控有反应的正向启动子元件失活。包含该元件的上游片段赋予LEU2 - lacZ融合基因cAMP负调控作用。对存在或不存在血红素的情况下、RAS2 +(高cAMP)和ras2突变体(低cAMP)菌株中以及在低温(23摄氏度)下生长的细胞和热激细胞(37摄氏度)中CTT1表达的Northern分析表明,只有当影响其表达的三个因素(由血红素发出的氧化应激信号、营养饥饿(低cAMP)和热应激)中的至少两个激活CTT1启动子时,CTT1才会被显著诱导。