Gui Lei, Liang Yong, Wei Dongsheng, Zheng Wen, Xing Laijun, Li Mingchun
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin 300071, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Aug;24(8):1348-53.
Microarray analysis revealed that the expression of ferric reductase (FRP1) can be regulated by the Riml01 protein. In order to find new transcriptional regulatory element in the promoter of FRP1, we analyzed the 1000 bp sequence upstream of ATG to find 2 potential Riml01p binding sites. We generated site-specific mutations in each of the two sites and fused these mutated promoters to LacZ. Then the promoter-LacZ fusion construct was recombinant into wild type and riml01-/- strains for beta-galactosidase assay. The results revealed that the FRP1 was up-regulated in alkaline pH and this was caused by iron starvation. The -650 site, not the -160 site, had an important role in FRP1 Riml01p-dependent expression. We conclude that Riml01p may interact with the -650 binding site of the promoter to regulate the FRP1 expression.
微阵列分析显示,铁还原酶(FRP1)的表达可受Riml01蛋白调控。为了在FRP1启动子中找到新的转录调控元件,我们分析了ATG上游1000 bp的序列,以找到2个潜在的Riml01p结合位点。我们在这两个位点分别产生了位点特异性突变,并将这些突变的启动子与LacZ融合。然后将启动子-LacZ融合构建体重组到野生型和riml01 -/-菌株中进行β-半乳糖苷酶测定。结果显示,FRP1在碱性pH条件下上调,这是由铁饥饿引起的。-650位点而非-160位点在FRP1依赖Riml01p的表达中起重要作用。我们得出结论,Riml01p可能与启动子的-650结合位点相互作用以调控FRP1的表达。