García de Viedma D, Giraldo R, Ruiz-Echevarría M J, Lurz R, Díaz-Orejas R
Centro de Investigaciones Biológicas, C.S.I.C. Velázquez, Madrid, Spain.
J Mol Biol. 1995 Mar 24;247(2):211-23. doi: 10.1006/jmbi.1994.0134.
Transcription of the repA gene of the Pseudomonas plasmid pPS10 is initiated from a sigma 70 type promoter located 81 bp upstream from the repA gene, extends through the repA gene and the adjacent open reading frame, and ends 1114 nucleotides downstream. The repA promoter is repressed by interactions of the RepA protein with a region of 44 bp that extends from the -10 box of the promoter to the dnaA box of the origin of replication. The core of the repA operator region is formed by two in-phase invertedly repeated sequences of 8 bp, S1 and S2, that flank the -35 box of the promoter, and that share homology with the internal sequences of the iterons present in the origin of replication. RepA enters at the operator region first by protein-DNA interactions and subsequently by protein-protein interactions. These sequential interactions lead to the formation of high, medium and low-mobility electrophoretic complexes. Formation of the high-order complexes seems to be important for an efficient repression of the promoter. Interactions of RepA with the repA promoter region (repPO) occur more efficiently than with the origin of replication.
假单胞菌质粒pPS10的repA基因转录起始于位于repA基因上游81 bp处的一个σ70型启动子,延伸穿过repA基因和相邻的开放阅读框,并在下游1114个核苷酸处终止。repA启动子通过RepA蛋白与一个44 bp区域的相互作用而受到抑制,该区域从启动子的-10框延伸至复制起点的dnaA框。repA操纵区的核心由两个8 bp的同相位反向重复序列S1和S2组成,它们位于启动子的-35框两侧,并且与复制起点中存在的迭代子的内部序列具有同源性。RepA首先通过蛋白质-DNA相互作用进入操纵区,随后通过蛋白质-蛋白质相互作用。这些顺序相互作用导致形成高、中、低迁移率的电泳复合物。高阶复合物的形成似乎对启动子的有效抑制很重要。RepA与repA启动子区域(repPO)的相互作用比与复制起点的相互作用更有效。