Pereira G G, Hollenberg C P
Institut für Mikrobiologie, Heinrich-Heine-Universität Düsseldorf, Germany.
Eur J Biochem. 1996 May 15;238(1):181-91. doi: 10.1111/j.1432-1033.1996.0181q.x.
The Hansenula polymorpha MOX gene encodes a peroxisomal enzyme that catalyzes the first step of the highly specialized methanol metabolism. MOX is strongly transcribed in cells growing in methanol and completely repressed in glucose. We show here that the MOX promoter confers a glucose-repressible expression upon a lacZ reporter gene in Saccharomyces cerevisiae, an unrelated yeast species that lacks the methanol metabolism. Repression was mediated by a 200-bp region of the MOX promoter, termed MOX-B, and was counteracted by Adr1p, a transcription factor involved in the derepression of S. cerevisiae genes encoding peroxisomal proteins, the class to which MOX belongs. Binding of Adr1p to MOX-B was demonstrated by gel retardation and DNaseI-footprinting, and Adr1p was shown to interact with a DNA region containing only a half of the putative Adr1p consensus binding site. Our findings suggest that Adr1p is a conserved regulator for genes encoding peroxisomal proteins at least in other yeast species, and that its interaction with the DNA is dependent on the promoter context.
多形汉逊酵母的MOX基因编码一种过氧化物酶体酶,该酶催化高度特化的甲醇代谢的第一步。MOX在以甲醇为生长底物的细胞中强烈转录,而在葡萄糖存在时完全被抑制。我们在此表明,MOX启动子可使酿酒酵母(一种缺乏甲醇代谢的无关酵母物种)中的lacZ报告基因实现葡萄糖可抑制表达。抑制作用由MOX启动子的一个200bp区域介导,该区域称为MOX-B,并且被Adr1p抵消,Adr1p是一种转录因子,参与酿酒酵母中编码过氧化物酶体蛋白(MOX所属类别)的基因的去抑制过程。通过凝胶阻滞和DNaseI足迹实验证明了Adr1p与MOX-B的结合,并且显示Adr1p与仅包含假定的Adr1p共有结合位点一半的DNA区域相互作用。我们的研究结果表明,Adr1p至少在其他酵母物种中是编码过氧化物酶体蛋白的基因的保守调节因子,并且其与DNA的相互作用取决于启动子背景。