Department of Chemistry, Hyogo College of Medicine, Nishinomiya, Japan.
FEMS Microbiol Lett. 2012 May;330(2):140-7. doi: 10.1111/j.1574-6968.2012.02543.x. Epub 2012 Apr 2.
In Saccharomyces cerevisiae, genes involved in thiamin pyrophosphate (TPP) synthesis (THI genes) and the pyruvate decarboxylase structural gene PDC5 are transcriptionally induced in response to thiamin starvation. Three positive regulatory factors (Thi2p, Thi3p, and Pdc2p) are involved in the expression of THI genes, whereas only Pdc2p is required for the expression of PDC5. Thi2p and Pdc2p serve as transcriptional activators and each factor can interact with Thi3p. The target consensus DNA sequence of Thi2p has been deduced. When TPP is not bound to Thi3p, the interactions between the regulatory factors are increased and THI gene expression is upregulated. In this study, we demonstrated that Pdc2p interacts with the upstream region of THI genes and PDC5. The association of Pdc2p or Thi2p with THI gene promoters was enhanced by thiamin starvation, suggesting that Pdc2p and Thi2p assist each other in their recruitment to the THI promoters via interaction with Thi3p. It is highly likely that, under thiamin-deprived conditions, a ternary Thi2p/Thi3p/Pdc2p complex is formed and transactivates THI genes in yeast cells. On the other hand, the association of Pdc2p with PDC5 was unaffected by thiamin. We also identified a DNA element in the upstream region of PDC5, which can bind to Pdc2p and is required for the expression of PDC5.
在酿酒酵母中,参与硫胺素焦磷酸(TPP)合成(THI 基因)和丙酮酸脱羧酶结构基因 PDC5 的基因在硫胺素饥饿时被转录诱导。三个正调控因子(Thi2p、Thi3p 和 Pdc2p)参与 THI 基因的表达,而只有 Pdc2p 是 PDC5 表达所必需的。Thi2p 和 Pdc2p 作为转录激活因子,每个因子都可以与 Thi3p 相互作用。已经推导出 Thi2p 的靶 DNA 序列的保守序列。当 TPP 未与 Thi3p 结合时,调节因子之间的相互作用增加,THI 基因表达上调。在这项研究中,我们证明了 Pdc2p 与 THI 基因和 PDC5 的上游区域相互作用。Thi3p 饥饿增强了 Pdc2p 或 Thi2p 与 THI 基因启动子的结合,表明 Pdc2p 和 Thi2p 通过与 Thi3p 的相互作用相互协助招募到 THI 启动子。在缺乏硫胺素的条件下,很可能形成三元 Thi2p/Thi3p/Pdc2p 复合物,从而在酵母细胞中转激活 THI 基因。另一方面,Pdc2p 与 PDC5 的结合不受硫胺素的影响。我们还鉴定了 PDC5 上游区域的一个 DNA 元件,该元件可以与 Pdc2p 结合,是 PDC5 表达所必需的。