Suzuki A, Tanifuji S, Komeda Y, Kato A
Division of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, Japan.
Plant Cell Physiol. 1995 Sep;36(6):1051-8. doi: 10.1093/oxfordjournals.pcp.a078847.
We previously described the preparation of a protein fraction that bound to the promoter region of the rRNA gene of Vicia faba. We now describe two additional protein fractions that interact with this DNA element. The three protein fractions have similar binding properties with the exception of their binding stabilities. DNase I footprinting experiments revealed that these protein fractions bind to a region that is located about 60 bp upstream from the site of initiation of transcription. From methylation interference experiments and gel-retardation assays with mutated oligonucleotides, it was evident that the protein fractions required the same residues for the formation of stable DNA-protein(s) complexes. However, the three protein fractions were chromatographically distinguishable and the stability of their binding to DNA was different. Although the functions of these fractions are still unknown, the fact that several protein fractions can bind to a narrow region near the site of initiation of transcription is of considerable interest in terms of the mechanism of transcription of the rRNA gene.
我们之前描述了一种与蚕豆rRNA基因启动子区域结合的蛋白质组分的制备方法。现在我们描述另外两种与该DNA元件相互作用的蛋白质组分。这三种蛋白质组分除了结合稳定性外,具有相似的结合特性。DNase I足迹实验表明,这些蛋白质组分结合在转录起始位点上游约60 bp的区域。从甲基化干扰实验以及用突变寡核苷酸进行的凝胶阻滞分析可知,这些蛋白质组分形成稳定的DNA-蛋白质复合物需要相同的残基。然而,这三种蛋白质组分在色谱上是可区分的,并且它们与DNA结合的稳定性不同。尽管这些组分的功能仍然未知,但就rRNA基因的转录机制而言,几种蛋白质组分能够结合在转录起始位点附近的狭窄区域这一事实相当令人感兴趣。