Braun R E, O'Day K, Wright A
Cell. 1985 Jan;40(1):159-69. doi: 10.1016/0092-8674(85)90319-8.
The dnaA gene in E. coli K-12 is required for the initiation of DNA replication. Although the specific function of the dnaA protein is unknown, it has been suggested that it is a regulator of the frequency of initiation. In this paper we report that the expression of both a dnaA-lacZ translational fusion and a dnaA-trpA-lacZ transcriptional fusion in vivo are sensitive to changes in the level of functional dnaA protein. Overproduction of the dnaA gene product leads to a reduction in expression from both fusions while introduction of dnaA- alleles results in an increased expression. Results from a deletion analysis of the dnaA promoter/regulatory region suggest that both dnaA promoters are regulated by the dnaA gene product and that a site between the two promoters is responsible for the regulation. DNAase protection experiments showed that the dnaA protein binds to DNA in the region of the two dnaA promoters. Our results indicate that the dnaA gene product regulates its own synthesis by inhibiting transcription from both of its promoters.
大肠杆菌K-12中的dnaA基因是DNA复制起始所必需的。虽然dnaA蛋白的具体功能尚不清楚,但有人认为它是起始频率的调节因子。在本文中,我们报道了体内dnaA-lacZ翻译融合和dnaA-trpA-lacZ转录融合的表达对功能性dnaA蛋白水平的变化敏感。dnaA基因产物的过量产生导致两种融合体的表达降低,而引入dnaA等位基因则导致表达增加。对dnaA启动子/调控区域的缺失分析结果表明,两个dnaA启动子均受dnaA基因产物调控,且两个启动子之间的一个位点负责这种调控。DNA酶保护实验表明,dnaA蛋白与两个dnaA启动子区域的DNA结合。我们的结果表明,dnaA基因产物通过抑制其两个启动子的转录来调节自身的合成。