Smith S D, Oriahi E, Yang-Yen H F, Xie W Q, Chen C, Rothblum L I
Weis Center for Research, Geisinger Clinic, Danville, PA 17822.
Nucleic Acids Res. 1990 Apr 11;18(7):1677-85. doi: 10.1093/nar/18.7.1677.
The spacer promoter of the rat rDNA repeat consists of two functional domains: a core (proximal) element that is sufficient for transcription in vitro, and an upstream (distal) promoter element that increases the efficiency of transcription. Two of the transcription factors that interact with the 45S promoter also interact with the spacer promoter. Rat SL-1, is required for transcription of the spacer promoter by heterologous extracts, e.g. human, and rat SF-1 is required for efficient transcription in vitro. Order-of-addition experiments demonstrated that the preinitiation complex formed by these factors on the spacer promoter is not as stable as the complex formed on the 45S promoter. DNase 1 footprinting experiments demonstrated binding sites for rat SL-1 and SF-1 on the distal element of the spacer promoter. The topology of the domains of the spacer promoter may explain both the reduced stability of the preinitiation complex formed on that promoter and the lower efficiency of transcription of that promoter when compared to the 45S promoter.
大鼠核糖体DNA(rDNA)重复序列的间隔区启动子由两个功能域组成:一个核心(近端)元件,其足以在体外进行转录,以及一个上游(远端)启动子元件,可提高转录效率。与45S启动子相互作用的两种转录因子也与间隔区启动子相互作用。大鼠SL-1是异源提取物(如人源提取物)转录间隔区启动子所必需的,而大鼠SF-1是体外高效转录所必需的。添加顺序实验表明,这些因子在间隔区启动子上形成的预起始复合物不如在45S启动子上形成的复合物稳定。DNA酶I足迹实验证明了大鼠SL-1和SF-1在间隔区启动子远端元件上的结合位点。与45S启动子相比,间隔区启动子结构域的拓扑结构可能解释了在该启动子上形成的预起始复合物稳定性降低以及该启动子转录效率较低的原因。