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首页> 外文期刊>Yeast >Analysis of the hypoxia-induced ADH2 promoter of the respiratory yeast Pichia stipitis reveals a new mechanism for sensing of oxygen limitation in yeast
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Analysis of the hypoxia-induced ADH2 promoter of the respiratory yeast Pichia stipitis reveals a new mechanism for sensing of oxygen limitation in yeast

机译：对呼吸道酵母毕赤酵母低氧诱导的ADH2启动子的分析揭示了一种新的机制，可检测酵母中的氧极限

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摘要

We introduced a reporter gene system into Pichia stipitis using the gene for the artificial green fluorescent protein (GFP), variant yEGFP. This system was used to analyse hypoxia-dependent PsADH2 regulation. Reporter gene activity was only found under oxygen limitation on a fermentable carbon source. The promoter was not induced by oxygen limitation in the Crabtree-positive yeast Saccharomyces cerevisiae. Promoter deletions revealed that a region of 15 bp contained the essential site for hypoxic induction. This motif was different from the known hypoxia response elements of S. cerevisiae but showed some similarity to the mammalian HIF-1 binding site. Electrophoretic mobility shift assays demonstrated specific protein binding to this region under oxygen limitation. Similar to the S. cerevisiae heme sensor system, the promoter was induced by Co2+. Cyanide was not able to mimic the effect of oxygen limitation. The activation mechanism of PsADH2 also, in this respect, has similarities to the mammalian HIF-1 system, which is inducible by Co2+ but not by cyanide. Thus, the very first promoter analysis in P. stipitis revealed a hitherto unknown mechanism of oxygen sensing in yeast.
机译：我们使用人工绿色荧光蛋白（GFP）变体yEGFP的基因将报告基因系统引入到毕赤酵母中。该系统用于分析缺氧依赖性的PsADH2调节。仅在可发酵碳源的氧气限制下才发现报告基因的活性。 Crabtree阳性酵母酿酒酵母中的氧限制没有诱导启动子。启动子缺失显示15 bp的区域含有缺氧诱导的必要位点。该基序不同于酿酒酵母已知的缺氧反应元件，但显示出与哺乳动物HIF-1结合位点的相似性。电泳迁移率变动分析表明在氧气限制下，特定蛋白质与该区域结合。类似于酿酒酵母血红素传感器系统，启动子是由Co2 +诱导的。氰化物不能模拟氧气限制的作用。在这方面，PsADH2的激活机制也与哺乳动物的HIF-1系统相似，该系统可被Co2 +诱导但不能被氰化物诱导。因此，对树干毕赤酵母的第一个启动子分析揭示了酵母中氧感测的迄今未知的机制。

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