Wang L, Helmann J D, Winans S C
Section of Microbiology, Cornell University, Ithaca, New York 14853.
Cell. 1992 May 15;69(4):659-67. doi: 10.1016/0092-8674(92)90229-6.
Octopine is released from crown gall tumors as a nutrient source and a signal molecule for the plant pathogen Agrobacterium tumefaciens. Some or all octopine-inducible genes are regulated by a protein called OccR. Primer extension analysis showed that OccR protein represses the occR gene and both represses and activates the occQ operon, which is divergently transcribed from occR. These promoters initiate transcription 46 bp apart. This regulatory system was reconstituted in vitro using purified OccR protein and Escherichia coli RNA polymerase. OccR binds with high affinity to a single site overlapping these promoters. Octopine shortens the DNAase I footprint of OccR and increases the gel mobility of OccR-DNA complexes by relaxing an OccR-incited DNA bend.
章鱼碱作为一种营养源和植物病原菌根癌土壤杆菌的信号分子从冠瘿瘤中释放出来。一些或所有章鱼碱诱导型基因受一种名为OccR的蛋白质调控。引物延伸分析表明,OccR蛋白抑制occR基因,同时抑制并激活从occR反向转录的occQ操纵子。这些启动子起始转录的位置相距46 bp。利用纯化的OccR蛋白和大肠杆菌RNA聚合酶在体外重建了这个调控系统。OccR以高亲和力结合到一个与这些启动子重叠的单一位点。章鱼碱缩短了OccR的DNA酶I足迹,并通过放松OccR引起的DNA弯曲增加了OccR-DNA复合物的凝胶迁移率。