Guilfoile P G, Hutchinson C R
School of Pharmacy, University of Wisconsin, Madison 53706.
J Bacteriol. 1992 Jun;174(11):3659-66. doi: 10.1128/jb.174.11.3659-3666.1992.
Preliminary evidence has been presented by Guilfoile and Hutchinson (J. Bacteriol. 174:3651-3658, 1992) suggesting that the Streptomyces glaucescens TcmR protein is a transcriptional repressor. Here, we extend that work by showing that transcription of the S. glaucescens tcmA gene is inducible by tetracenomycin C and that inactivation of the tcmR gene results in constitutive transcription of the tcmA gene. Gel retardation studies show that the TcmR protein binds to the tcmA-tcmR intergenic region in vitro and that this binding is inhibited by tetracenomycin C. Footprinting experiments demonstrate that the TcmR protein binds to an operator region that encompasses both the tcmA and the tcmR promoters. This genetic and biochemical evidence strongly supports the model of the TcmR protein acting as a repressor in inhibiting transcription of both the tcmA and the tcmR genes, in much the same way that TetR from Tn10 inhibits transcription of tetA and tetR.
吉尔福伊尔和哈钦森(《细菌学杂志》174:3651 - 3658,1992年)已给出初步证据,表明青灰链霉菌TcmR蛋白是一种转录阻遏物。在此,我们拓展了该研究,表明青灰链霉菌tcmA基因的转录可被四环素霉素C诱导,且tcmR基因的失活会导致tcmA基因的组成型转录。凝胶阻滞研究表明,TcmR蛋白在体外与tcmA - tcmR基因间区域结合,且这种结合会被四环素霉素C抑制。足迹实验证明,TcmR蛋白与一个包含tcmA和tcmR启动子的操纵区结合。这些遗传学和生物化学证据有力地支持了TcmR蛋白作为阻遏物抑制tcmA和tcmR基因转录的模型,其方式与Tn10的TetR抑制tetA和tetR转录的方式非常相似。