Schmid A, Fascher K D, Hörz W
Institute for Physiological Chemistry, Universität München, Germany.
Cell. 1992 Nov 27;71(5):853-64. doi: 10.1016/0092-8674(92)90560-y.
Activation of the PHO5 gene in S. cerevisiae by phosphate starvation was previously shown to be accompanied by the disappearance of four positioned nucleosomes from the promoter. To investigate the mechanism, we replaced the PHO80 gene, a negative regulator of PHO5, by a temperature-sensitive allele. As a consequence, PHO5 can be activated in the presence of phosphate by a temperature shift from 24 degrees C to 37 degrees C. Under these conditions, the promoter undergoes the same chromatin transition as in phosphate-starved cells. Disruption of the nucleosomes by the temperature shift also occurs when DNA replication is prevented. Nucleosomes re-form when the temperature is shifted from 37 degrees C back to 24 degrees C in nondividing cells. Glucose is required for the disruption of the nucleosomes during the temperature upshift, not for their re-formation during the temperature downshift. These experiments prove that DNA replication is not required for the transition between the nucleosomal and the non-nucleosomal state at the PHO5 promoter.
先前的研究表明,酿酒酵母中PHO5基因在磷酸盐饥饿时的激活伴随着启动子区域四个定位核小体的消失。为了探究其机制,我们用一个温度敏感等位基因替换了PHO5的负调控因子PHO80基因。结果,通过将温度从24摄氏度转变为37摄氏度,PHO5在磷酸盐存在的情况下也能够被激活。在这些条件下,启动子经历了与磷酸盐饥饿细胞中相同的染色质转变。当DNA复制被阻止时,温度转变导致的核小体破坏依然会发生。在非分裂细胞中,当温度从37摄氏度变回24摄氏度时,核小体会重新形成。在温度上升过程中,核小体的破坏需要葡萄糖,但在温度下降过程中核小体的重新形成则不需要葡萄糖。这些实验证明,PHO5启动子处核小体状态与非核小体状态之间的转变并不需要DNA复制。