Benner-Luger D, Boos W
Department of Biology, University of Konstanz, Federal Republic of Germany.
Mol Gen Genet. 1988 Nov;214(3):579-87. doi: 10.1007/BF00330498.
The mglB gene of Salmonella typhimurium LT2 coding for the galactose-binding protein (GBP) was sequenced. We compared the deduced amino acid sequence with the GBP sequence of Escherichia coli K 12. The mature proteins differ in only 19 of 309 amino acid residues, corresponding to 94% homology. Analysis of the mglB control region by promoter-probe vectors revealed that two promoters, P1 and P2, constitute the mgl control region (Pmgl). P1 and P2 function in a synergistic way. P1 is the main promoter of the operon; its activity is 20 times the activity of P2. Both promoters are activated by the cyclic adenosine monophosphate catabolite activator protein (cAMP/CAP) complex. While P1 is inactive in the absence of the cAMP/CAP complex, there is residual activity of P2 under these conditions. Studies on the inducibility of the mglBAEC operon using multicopy plasmid promoter-probe vectors were hampered by the titration of the mgl repressor resulting in a partially constitutive expression of the mgl operon. The results indicate that only P1 is responding to induction by D-fucose. A weak promoter, PD, within the P1 region but divergent to it was found. PD is neither stimulated by the cAMP/CAP complex nor by D-fucose. We cloned the gene located downstream to PD and found it to strongly repress the expression of the mgl operon. We termed this gene mglD. The presence of D-fucose abolished the repression caused by the plasmid-encoded mglD gene product.
对鼠伤寒沙门氏菌LT2编码半乳糖结合蛋白(GBP)的mglB基因进行了测序。我们将推导的氨基酸序列与大肠杆菌K12的GBP序列进行了比较。成熟蛋白在309个氨基酸残基中仅有19个不同,对应94%的同源性。通过启动子探针载体对mglB调控区进行分析发现,两个启动子P1和P2构成了mgl调控区(Pmgl)。P1和P2以协同方式发挥作用。P1是操纵子的主要启动子;其活性是P2活性的20倍。两个启动子均由环磷酸腺苷分解代谢物激活蛋白(cAMP/CAP)复合物激活。虽然在没有cAMP/CAP复合物的情况下P1无活性,但在这些条件下P2仍有残余活性。使用多拷贝质粒启动子探针载体对mglBAEC操纵子的诱导性进行的研究受到mgl阻遏物滴定的阻碍,导致mgl操纵子部分组成型表达。结果表明,只有P1对D-岩藻糖的诱导有反应。在P1区域内但与之方向相反发现了一个弱启动子PD。PD既不受cAMP/CAP复合物的刺激,也不受D-岩藻糖的刺激。我们克隆了位于PD下游的基因,发现它强烈抑制mgl操纵子的表达。我们将该基因命名为mglD。D-岩藻糖的存在消除了由质粒编码的mglD基因产物引起的抑制作用。