Nevado J, Gaudreau L, Adam M, Ptashne M
Molecular Biology Program, Sloan-Kettering Institute, New York, NY 10021, USA.
Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):2674-7. doi: 10.1073/pnas.96.6.2674.
We show that the typical "nonclassical" activator, which comprises a fusion protein bearing a component of the transcriptional machinery fused to a DNA-binding domain, activates transcription in mammalian cells only weakly when tested with an array of promoters. However, as found in analogous "artificial recruitment" experiments performed in yeast, these activators work synergistically with "classical" activators. The effect of the classical activator in such experiments requires that it be tethered to DNA, a requirement that cannot be overcome by expression of that classical activator at high levels. The effect of the one nonclassical activator that does elicit significant levels of transcription when working alone (i.e., that bearing TATA box-binding protein) is strongly influenced by promoter architecture. The results, consistent with those of analogous experiments in yeast [see the accompanying paper: Gaudreau, L., Keaveney, M., Nevado, J., Zaman, Z., Bryant, G. O., Struhl, K. & Ptashne, M. (1999) Proc. Natl. Acad. Sci. USA 96, 2668-2673], suggest that classical activators, presumably by virtue of their abilities to interact with multiple targets, have a functional flexibility that nonclassical activators lack.
我们发现,典型的“非经典”激活因子,即一种转录机制的组成部分与DNA结合结构域融合而成的融合蛋白,在用一系列启动子进行测试时,在哺乳动物细胞中仅能微弱地激活转录。然而,正如在酵母中进行的类似“人工招募”实验所发现的那样,这些激活因子能与“经典”激活因子协同发挥作用。在这类实验中,经典激活因子发挥作用需要将其与DNA相连,即使高水平表达该经典激活因子也无法克服这一要求。有一种非经典激活因子单独作用时能引发显著水平的转录(即带有TATA盒结合蛋白的激活因子),其作用受到启动子结构的强烈影响。这些结果与酵母中类似实验的结果一致[见随附论文:Gaudreau, L., Keaveney, M., Nevado, J., Zaman, Z., Bryant, G. O., Struhl, K. & Ptashne, M. (1999) Proc. Natl. Acad. Sci. USA 96, 2668 - 2673],表明经典激活因子可能凭借其与多个靶点相互作用的能力,具有非经典激活因子所缺乏的功能灵活性。