Raitt D C, Johnson A L, Erkine A M, Makino K, Morgan B, Gross D S, Johnston L H
Division of Yeast Genetics, National Institute for Medical Research, The Ridgeway, London NW7 1AA, United Kingdom.
Mol Biol Cell. 2000 Jul;11(7):2335-47. doi: 10.1091/mbc.11.7.2335.
The Skn7 response regulator has previously been shown to play a role in the induction of stress-responsive genes in yeast, e.g., in the induction of the thioredoxin gene in response to hydrogen peroxide. The yeast Heat Shock Factor, Hsf1, is central to the induction of another set of stress-inducible genes, namely the heat shock genes. These two regulatory trans-activators, Hsf1 and Skn7, share certain structural homologies, particularly in their DNA-binding domains and the presence of adjacent regions of coiled-coil structure, which are known to mediate protein-protein interactions. Here, we provide evidence that Hsf1 and Skn7 interact in vitro and in vivo and we show that Skn7 can bind to the same regulatory sequences as Hsf1, namely heat shock elements. Furthermore, we demonstrate that a strain deleted for the SKN7 gene and containing a temperature-sensitive mutation in Hsf1 is hypersensitive to oxidative stress. Our data suggest that Skn7 and Hsf1 cooperate to achieve maximal induction of heat shock genes in response specifically to oxidative stress. We further show that, like Hsf1, Skn7 can interact with itself and is localized to the nucleus under normal growth conditions as well as during oxidative stress.
此前已表明,Skn7反应调节因子在酵母中应激反应基因的诱导过程中发挥作用,例如在过氧化氢诱导硫氧还蛋白基因的过程中。酵母热休克因子Hsf1对于另一组应激诱导基因(即热休克基因)的诱导至关重要。这两种调节性反式激活因子Hsf1和Skn7具有某些结构同源性,特别是在它们的DNA结合结构域以及存在已知介导蛋白质-蛋白质相互作用的相邻卷曲螺旋结构区域方面。在这里,我们提供证据表明Hsf1和Skn7在体外和体内相互作用,并且我们表明Skn7可以与Hsf1结合相同的调控序列,即热休克元件。此外,我们证明缺失SKN7基因且在Hsf1中含有温度敏感突变的菌株对氧化应激高度敏感。我们的数据表明,Skn7和Hsf1协同作用,以实现热休克基因在对氧化应激的特异性反应中的最大诱导。我们进一步表明,与Hsf1一样,Skn7可以自身相互作用,并且在正常生长条件下以及氧化应激期间定位于细胞核。