Pierce J, Fee B E, Toohey M G, Peterson D O
Department of Biochemistry and Biophysics, Texas A&M University, College Station 77843-2128.
J Virol. 1993 Jan;67(1):415-24. doi: 10.1128/JVI.67.1.415-424.1993.
Transcription from the promoter of mouse mammary tumor virus is subject to both positive and negative control by cellular factors, and proviral promoter elements that mediate a basal level of transcription must in some way respond to these cellular regulatory signals. Several such elements, including a TATA box, a region containing three octamer-related sequences, and a binding site for nuclear factor 1, have been previously defined. Additional promoter mutations have allowed a fourth basal promoter element to be identified near the transcription initiation site between +2 and +10. Sequence alterations within this element affect transcription both in vivo and in vitro. Gel electrophoresis mobility shift and DNase I footprinting assays define a nuclear protein, termed initiation site-binding protein, that specifically recognizes this region of the promoter. Optimal levels of transcription from the mouse mammary tumor virus promoter require initiation site-binding protein, as demonstrated by a correlation between protein affinity and transcriptional activity and by specific inhibition of transcription in vitro by an oligonucleotide capable of titrating the protein from transcriptionally active fractions.
小鼠乳腺肿瘤病毒启动子的转录受到细胞因子的正调控和负调控,介导基础转录水平的前病毒启动子元件必须以某种方式响应这些细胞调节信号。此前已确定了几个这样的元件,包括一个TATA盒、一个包含三个八聚体相关序列的区域以及一个核因子1结合位点。额外的启动子突变使得在转录起始位点附近+2至+10之间鉴定出了第四个基础启动子元件。该元件内的序列改变在体内和体外均影响转录。凝胶电泳迁移率变动分析和DNase I足迹分析确定了一种核蛋白,称为起始位点结合蛋白,它能特异性识别启动子的这一区域。蛋白质亲和力与转录活性之间的相关性以及一种能够从转录活性组分中滴定该蛋白质的寡核苷酸在体外对转录的特异性抑制表明,小鼠乳腺肿瘤病毒启动子的最佳转录水平需要起始位点结合蛋白。