Hirota Kouji, Hoffman Charles S, Ohta Kunihiro
Genetic System Regulation Laboratory, RIKEN (The Institute of Physical and Chemical Research), Discovery Research Institute, Wako-shi, Saitama 351-0198, Japan.
Eukaryot Cell. 2006 Dec;5(12):1980-9. doi: 10.1128/EC.00272-06. Epub 2006 Oct 6.
The Schizosaccharomyces pombe global corepressors Tup11 and Tup12, which are orthologs of Saccharomyces cerevisiae Tup1, are involved in glucose-dependent transcriptional repression and chromatin alteration of the fbp1+ gene. The fbp1+ promoter contains two regulatory elements, UAS1 and UAS2, one of which (UAS2) serves as a binding site for two antagonizing C2H2 Zn finger transcription factors, the Rst2 activator and the Scr1 repressor. In this study, we analyzed the role of Tup proteins and Scr1 in chromatin remodeling at fbp1+ during glucose repression. We found that Scr1, cooperating with Tup11 and Tup12, functions to maintain the chromatin of the fbp1+ promoter in a transcriptionally inactive state under glucose-rich conditions. Consistent with this notion, Scr1 is quickly exported from the nucleus to the cytoplasm at the initial stage of derepression, immediately after glucose starvation, at which time Rst2 is known to be imported into the nucleus. In addition, chromatin immunoprecipitation assays revealed a switching of Scr1 to Rst2 bound at UAS2 during glucose derepression. On the other hand, Tup11 and Tup12 persist in the nucleus and bind to the fbp1+ promoter under both derepressed and repressed conditions. These observations suggest that Tup1-like proteins recruited to the fbp1+ promoter are controlled by either of two antagonizing C2H2 Zn finger proteins. We propose that the actions of Tup11 and Tup12 are regulated by reciprocal nuclear shuttling of the two antagonizing Zn finger proteins in response to the extracellular glucose concentration. This notion provides new insights into the molecular mechanisms of the Tup family corepressors in gene regulation.
粟酒裂殖酵母全局共抑制因子Tup11和Tup12是酿酒酵母Tup1的直系同源物,参与葡萄糖依赖性转录抑制和fbp1 +基因的染色质改变。fbp1 +启动子包含两个调控元件UAS1和UAS2调控元件,其中一个(UAS2)作为两种拮抗性C2H2锌指转录因子Rst2激活剂和Scr1阻遏物的结合位点。在本研究中,我们分析了Tup蛋白和Scr1在葡萄糖抑制期间fbp1 +染色质重塑中的作用。我们发现,Scr1与Tup11和Tup12协同作用,在富含葡萄糖的条件下,将fbp1 +启动子的染色质维持在转录非活性状态。与此概念一致,在去抑制初期,即在葡萄糖饥饿后,Scr1迅速从细胞核输出到细胞质,此时已知Rst2被导入细胞核。此外,染色质免疫沉淀试验显示,在葡萄糖去抑制过程中,Scr1与结合在UAS2上的Rst2发生了转换。另一方面,Tup11和Tup12在细胞核中持续存在,并在去抑制和抑制条件下均与fbp1 +启动子结合。这些观察结果表明,招募到fbp1 +启动子的Tup1样蛋白受两种拮抗性C2H2锌指蛋白之一的控制。我们提出,Tup11和Tup12的作用受两种拮抗性锌指蛋白响应细胞外葡萄糖浓度的相互核穿梭调节。这一概念为Tup家族共抑制因子在基因调控中的分子机制提供了新的见解。