Furuya K, Hutchinson C R
School of Pharmacy, University of Wisconsin, Madison 53706, USA.
FEMS Microbiol Lett. 1998 Nov 15;168(2):243-9. doi: 10.1111/j.1574-6968.1998.tb13280.x.
DrrC, a daunorubicin resistance protein with a strong sequence similarity to the UvrA protein involved in excision repair of DNA, is induced by daunorubicin in Streptomyces peucetius and behaves like an ATP-dependent, DNA binding protein in vitro. The refolded protein obtained from expression of the drrC gene in Escherichia coli was used to conduct gel retardation assays. DrrC bound a DNA segment containing the promoter region of a daunorubicin production gene only in the presence of ATP and daunorubicin. This result suggests that DrrC is a novel type of drug self-resistance protein with DNA binding properties like those of UvrA. Western blotting analysis with a polyclonal antiserum generated against His-tagged DrrC showed that the appearance of DrrC in S. peucetius is coincident with the onset of daunorubicin production and that the drrC gene is induced by daunorubicin. These data also showed that the DnrN and DnrI regulatory proteins are required for drrC expression. The level of DrrA, another daunorubicin resistance protein that resembles ATP-dependent bacterial antiporters, was regulated in the same way as that of DrrC.
DrrC是一种柔红霉素抗性蛋白，与参与DNA切除修复的UvrA蛋白具有很强的序列相似性，在变铅青链霉菌中由柔红霉素诱导产生，在体外表现为一种依赖ATP的DNA结合蛋白。利用在大肠杆菌中表达drrC基因获得的重折叠蛋白进行凝胶阻滞分析。DrrC仅在ATP和柔红霉素存在的情况下结合包含柔红霉素生产基因启动子区域的DNA片段。这一结果表明，DrrC是一种新型的具有DNA结合特性的药物自身抗性蛋白，类似于UvrA。用针对His标签的DrrC产生的多克隆抗血清进行的蛋白质印迹分析表明，DrrC在变铅青链霉菌中的出现与柔红霉素产生的开始相一致，并且drrC基因由柔红霉素诱导。这些数据还表明，DnrN和DnrI调节蛋白是drrC表达所必需的。另一种柔红霉素抗性蛋白DrrA，类似于依赖ATP的细菌反向转运体，其水平与DrrC的调节方式相同。
