Gilmour D S, Lis J T
Mol Cell Biol. 1986 Nov;6(11):3984-9. doi: 10.1128/mcb.6.11.3984-3989.1986.
By using a protein-DNA cross-linking method (D. S. Gilmour and J. T. Lis, Mol. Cell. Biol. 5:2009-2018, 1985), we examined the in vivo distribution of RNA polymerase II on the hsp70 heat shock gene in Drosophila melanogaster Schneider line 2 cells. In heat shock-induced cells, a high level of RNA polymerase II was detected on the entire gene, while in noninduced cells, the RNA polymerase II was confined to the 5' end of the hsp70 gene, predominantly between nucleotides -12 and +65 relative to the start of transcription. This association of RNA polymerase II was apparent whether the cross-linking was performed by a 10-min UV irradiation of chilled cells with mercury vapor lamps or by a 40-microsecond irradiation of cells with a high-energy xenon flash lamp. We hypothesize that RNA polymerase II has access to, and a high affinity for, the promoter region of this gene before induction, and this poised RNA polymerase II may be critical in the mechanism of transcription activation.
通过使用一种蛋白质-DNA交联方法(D. S. 吉尔摩和J. T. 利斯,《分子细胞生物学》5:2009 - 2018,1985年),我们检测了RNA聚合酶II在黑腹果蝇施耐德2号线细胞中hsp70热休克基因上的体内分布情况。在热休克诱导的细胞中,在整个基因上检测到高水平的RNA聚合酶II,而在未诱导的细胞中,RNA聚合酶II局限于hsp70基因的5'端,主要在相对于转录起始点的核苷酸-12至+65之间。无论交联是通过用汞蒸气灯对冷冻细胞进行10分钟紫外线照射,还是通过用高能氙闪光灯对细胞进行40微秒照射来进行,RNA聚合酶II的这种结合都是明显的。我们推测,RNA聚合酶II在诱导之前能够接近该基因的启动子区域并对其具有高亲和力,而这种处于就绪状态的RNA聚合酶II可能在转录激活机制中至关重要。