Buratowski S, Sopta M, Greenblatt J, Sharp P A
Whitehead Institute for Biomedical Research, Cambridge, MA 02142.
Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7509-13. doi: 10.1073/pnas.88.17.7509.
Proteins purified on the basis of their affinity for RNA polymerase II effectively substitute for previously defined transcription initiation factors. In two assays, formation of initiation complexes and transcription in vitro, the RNA polymerase II-associated proteins behaved identically to a fraction containing transcription factors IIE and IIF. Both fractions greatly stabilized the association of polymerase with the promoter and were required for the formation of complete initiation complexes. By using the DNA-cleaving reagent phenanthroline.copper in footprinting reactions, the RNA polymerase II-associated proteins were shown to be required for a DNA conformation change near the initiation site of the promoter. Based on similarity to the prokaryotic transcription complex, this conformation change is likely to represent a transition from a closed to an open complex.
基于对RNA聚合酶II的亲和性而纯化的蛋白质能够有效地替代先前定义的转录起始因子。在两项检测中,即起始复合物的形成和体外转录,与RNA聚合酶II相关的蛋白质的行为与含有转录因子IIE和IIF的组分完全相同。这两个组分都极大地稳定了聚合酶与启动子的结合,并且是形成完整起始复合物所必需的。通过在足迹反应中使用DNA切割试剂菲咯啉-铜,发现与RNA聚合酶II相关的蛋白质是启动子起始位点附近DNA构象变化所必需的。基于与原核转录复合物的相似性,这种构象变化可能代表从封闭复合物到开放复合物的转变。