Meng L M, Nygaard P
Institute of Biological Chemistry B, University of Copenhagen, Denmark.
Mol Microbiol. 1990 Dec;4(12):2187-92. doi: 10.1111/j.1365-2958.1990.tb00580.x.
Addition of purine compounds to the growth medium of Escherichia coli and Salmonella typhimurium causes repressed synthesis of the purine biosynthetic enzymes. The repression is mediated through a regulatory protein, PurR. To identify the co-repressor(s) of PurR, two approaches were used: (i) mutations were introduced into purine salvage genes and the effects of different purines on pur gene expression were determined; (ii) purine compounds which dictate the binding of the PurR protein to its operator DNA were resolved by gel retardation. Both the in vivo and the in vitro data indicated that guanine and hypoxanthine are co-repressors. The toxic purine analogues 6-mercaptopurine and 6-thioguanine also activated the binding of PurR to its operator DNA.
向大肠杆菌和鼠伤寒沙门氏菌的生长培养基中添加嘌呤化合物会导致嘌呤生物合成酶的合成受到抑制。这种抑制是通过一种调节蛋白PurR介导的。为了鉴定PurR的共阻遏物,采用了两种方法:(i)将突变引入嘌呤补救途径基因,并测定不同嘌呤对嘌呤基因表达的影响;(ii)通过凝胶阻滞法解析决定PurR蛋白与其操纵子DNA结合的嘌呤化合物。体内和体外数据均表明鸟嘌呤和次黄嘌呤是共阻遏物。有毒的嘌呤类似物6-巯基嘌呤和6-硫鸟嘌呤也能激活PurR与其操纵子DNA的结合。