Urbanowski M L, Stauffer L T, Plamann L S, Stauffer G V
J Bacteriol. 1987 Apr;169(4):1391-7. doi: 10.1128/jb.169.4.1391-1397.1987.
We isolated an Escherichia coli methionine auxotroph that displays a growth phenotype similar to that of known metF mutants but has elevated levels of 5,10-methylenetetrahydrofolate reductase, the metF gene product. Transduction analysis indicates that the mutant carries normal metE, metH, and metF genes; the phenotype is due to a single mutation, eliminating the possibility that the strain is a metE metH double mutant; and the new mutation is linked to the metE gene by P1 transduction. Plasmids carrying the Salmonella typhimurium metE gene and flanking regions complement the mutation, even when the plasmid-borne metE gene is inactivated. Enzyme assays show that the mutation results in a dramatic decrease in metE gene expression, a moderate decrease in metH gene expression, and a disruption of the metH-mediated vitamin B12 repression of the metE and metF genes. Our evidence suggests that the methionine auxotrophy caused by the new mutation is a result of insufficient production of both the vitamin B12-independent (metE) and vitamin B12-dependent (metH) transmethylase enzymes that are necessary for the synthesis of methionine from homocysteine. We propose that this mutation defines a positive regulatory gene, designated metR, whose product acts in trans to activate the metE and metH genes.
我们分离出了一株大肠杆菌甲硫氨酸营养缺陷型菌株,该菌株表现出与已知metF突变体相似的生长表型,但甲硫氨酸合成酶(metF基因产物)的水平有所升高。转导分析表明,该突变体携带正常的metE、metH和metF基因;其表型是由单一突变引起的,排除了该菌株是metE metH双突变体的可能性;并且通过P1转导发现新突变与metE基因连锁。携带鼠伤寒沙门氏菌metE基因及其侧翼区域的质粒能够互补该突变,即使质粒携带的metE基因失活时也是如此。酶活性测定表明,该突变导致metE基因表达显著下降,metH基因表达适度下降,并且破坏了metH介导的维生素B12对metE和metF基因的抑制作用。我们的证据表明,新突变引起的甲硫氨酸营养缺陷是由于同型半胱氨酸合成甲硫氨酸所需的维生素B12非依赖性(metE)和维生素B12依赖性(metH)转甲基酶产量不足所致。我们提出,这种突变定义了一个正向调节基因,命名为metR,其产物通过反式作用激活metE和metH基因。