Verwaal René, Arako Megumi, Kapur Rick, Verkleij Arie J, Verrips C Theo, Boonstra Johannes
Department of Molecular Cell Biology and the Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Yeast. 2004 Jul 15;21(9):747-57. doi: 10.1002/yea.1106.
Hexose transporter (Hxt) proteins transport glucose across the plasma membrane in the yeast Saccharomyces cerevisiae. Recently, we have shown that expression of HXT5 is regulated by the growth rate of the cells. Because gene expression is regulated by binding of specific transcription factors to regulatory elements in the promoters of genes, the presence of putative regulatory elements in the promoter of HXT5 was determined by computer-assisted analysis. This revealed the presence of two putative stress-responsive elements (STREs), one putative post-diauxic shift (PDS) element and two putative Hap2/3/4/5p (HAP) complex binding elements. The involvement of these elements was studied by using mutations in a HXT5 promoter-LacZ fusion construct. Growth during various conditions that result in low growth rates of yeast cells revealed that the STRE most proximal to the translation initiation site seemed to be involved in particular in regulation of HXT5 expression during growth at decreased growth rates. In addition, the HAP elements seemed to be required during growth on non-fermentable carbon sources. The PDS element and, to a lesser extent, the other STRE showed particular involvement in regulation of HXT5 expression during growth on ethanol. Finally, it was shown that the PKA pathway, which is known to be involved in expression of STRE-regulated genes, was also involved in regulation of HXT5 expression. A possible mechanism by which expression of HXT5 could be regulated by the transcriptional regulatory elements in the promoter is discussed.
己糖转运蛋白(Hxt)可将葡萄糖转运穿过酿酒酵母的质膜。最近,我们发现HXT5的表达受细胞生长速率的调控。由于基因表达是通过特定转录因子与基因启动子中的调控元件结合来调控的,因此通过计算机辅助分析确定了HXT5启动子中假定调控元件的存在。这揭示了两个假定的应激反应元件(STRE)、一个假定的二次生长转换后(PDS)元件以及两个假定的Hap2/3/4/5p(HAP)复合物结合元件。通过使用HXT5启动子-LacZ融合构建体中的突变来研究这些元件的作用。在导致酵母细胞生长速率较低的各种条件下生长的结果表明,最靠近翻译起始位点的STRE似乎特别参与了生长速率降低时HXT5表达的调控。此外,在以非发酵碳源生长期间,似乎需要HAP元件。PDS元件以及在较小程度上另一个STRE在乙醇生长期间对HXT5表达的调控中表现出特别的参与。最后,结果表明已知参与STRE调控基因表达的PKA途径也参与了HXT5表达的调控。本文讨论了启动子中的转录调控元件调控HXT5表达的可能机制。