Liang Yong, Gui Lei, Wei Dong-Sheng, Zheng Wen, Xing Lai-Jun, Li Ming-Chun
Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Department of Microbiology, Nankai University, Tianjin, China.
FEMS Yeast Res. 2009 Mar;9(2):270-7. doi: 10.1111/j.1567-1364.2008.00468.x. Epub 2008 Dec 9.
The opportunistic pathogen Candida albicans develops different systems to acquire essential nutrients such as iron. The Rim101 pathway controls gene expression to adapt to different environments through transcription factor Rim101p. Ferric reductase genes are important for iron acquisition, but further work should be carried out to show about how ferric reductase genes are regulated. In this study we demonstrate that ferric reductase FRP1 expression is upregulated by alkaline pH and iron-limited conditions. Moreover, promoter deletion identified that the upregulated elements lie in -822 to 437 and 263 to -124 and the downregulated elements in -437 to -263 using a beta-galactosidase reporter system. Using electrophoretic mobility shift assay we also found that Rim101p can bind directly with two putative 5'-CCAAGAA-3' sites at the -157 and -629 sites. Interestingly, site-specific mutations show that only the -629 site, not the -157 site, plays an important role in FRP1 expression under iron-limited conditions.
机会性致病真菌白色念珠菌会形成不同系统来获取铁等必需营养物质。Rim101途径通过转录因子Rim101p控制基因表达以适应不同环境。铁还原酶基因对铁的获取很重要,但关于铁还原酶基因如何被调控仍需进一步研究。在本研究中,我们证明碱性pH值和铁限制条件会上调铁还原酶FRP1的表达。此外,使用β-半乳糖苷酶报告系统进行启动子缺失实验发现,上调元件位于-822至437以及263至-124区域,而下调元件位于-437至-263区域。通过电泳迁移率变动分析,我们还发现Rim101p能直接与-157和-629位点处的两个假定的5'-CCAAGAA-3'位点结合。有趣的是,位点特异性突变表明,在铁限制条件下,只有-629位点而非-157位点对FRP1的表达起重要作用。