Ramseier T M, Bledig S, Michotey V, Feghali R, Saier M H
Department of Biology, University of California at San Diego, La Jolla 92093-0116, USA.
Mol Microbiol. 1995 Jun;16(6):1157-69. doi: 10.1111/j.1365-2958.1995.tb02339.x.
The Escherichia coli fructose repressor, FruR, is known to regulate expression of several genes concerned with carbon utilization. Using a previously derived consensus sequence for FruR binding, additional potential operators were identified and tested for FruR binding in DNA band migration retardation assays. Operators in the control regions of operons concerned with carbon metabolism bound FruR, while those in operons not concerned with carbon metabolism did not. In vivo assays with transcriptional lacZ fusions showed that FruR controls the expression of FruR operator-containing genes encoding key enzymes of virtually every major pathway of carbon metabolism. Moreover, a fruR null mutation altered the rates of utilization of at least 36 carbon sources. In general, oxidation rates for glycolytic substances were enhanced while those for gluconeogenic substances were depressed. Alignment of FruR operators revealed that the consensus sequence for FruR binding is the same for operons that are activated and repressed by FruR and permitted formulation of a revised FruR-binding consensus sequence. The reported observations indicate that FruR modulates the direction of carbon flow by transcriptional activation of genes encoding enzymes concerned with oxidative and gluconeogenic carbon flow and by repression of those concerned with fermentative carbon flow.
已知大肠杆菌果糖阻遏物FruR可调控多个与碳利用相关基因的表达。利用先前推导的FruR结合共有序列,在DNA条带迁移阻滞试验中鉴定并测试了其他潜在的操纵基因与FruR的结合情况。参与碳代谢的操纵子控制区域中的操纵基因能与FruR结合,而不参与碳代谢的操纵子中的操纵基因则不能。利用转录型lacZ融合体进行的体内试验表明,FruR可控制含FruR操纵基因的基因的表达,这些基因编码碳代谢几乎每一条主要途径的关键酶。此外,fruR基因敲除突变改变了至少36种碳源的利用速率。一般来说,糖酵解物质的氧化速率提高,而糖异生物质的氧化速率降低。FruR操纵基因的比对显示,FruR结合的共有序列对于受FruR激活和抑制的操纵子而言是相同的,从而得以制定修订后的FruR结合共有序列。所报道的观察结果表明,FruR通过转录激活编码与氧化和糖异生碳流相关酶的基因以及抑制与发酵碳流相关的基因,来调节碳流方向。