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酿酒酵母中HXT13、HXT15和HXT17基因的表达以及糖诱导的渗透胁迫对HXT1基因转录本的稳定作用。

Expression of the HXT13, HXT15 and HXT17 genes in Saccharomyces cerevisiae and stabilization of the HXT1 gene transcript by sugar-induced osmotic stress.

作者信息

Greatrix Bradley W, van Vuuren Hennie J J

机构信息

Wine Research Centre, The University of British Columbia, Suite 231#2205 East Mall, V6T 1Z4, Vancouver, Canada.

出版信息

Curr Genet. 2006 Apr;49(4):205-17. doi: 10.1007/s00294-005-0046-x. Epub 2006 Jan 6.

Abstract

Saccharomyces cerevisiae contains a family of 17 hexose transporter (HXT) genes; only nine have assigned functions, some of which are still poorly defined. Despite extensive efforts to characterize the hexose transporters, the expression of HXT6 and HXT8-17 remains an enigma. In nature, S. cerevisiae finds itself under extreme nutritional conditions including sugars in excess of 40% (w/v), depletion of nutrients and extremes of both temperature and pH. Using HXT promoter-lacZ fusions, we have identified novel conditions under which the HXT17 gene is expressed; HXT17 promoter activity is up-regulated in media containing raffinose and galactose at pH 7.7 versus pH 4.7. We demonstrated that HXT5, HXT13 and, to a lesser extent, HXT15 were all induced in the presence of non-fermentable carbon sources. HXT1 encodes a low-affinity transporter and in short-term osmotic shock experiments, HXT1 promoter activity was reduced when cells were exposed to media containing 40% glucose. However, we found that the HXT1 mRNA transcript was stabilized under conditions of osmotic stress. Furthermore, the stabilization of HXT1 mRNA does not appear to be gene specific because 30 min after transcriptional arrest there is a fourfold more mRNA in osmotically stressed versus non-stressed yeast cells. A large portion of S. cerevisiae mRNA molecules may, therefore, have a decreased rate of turnover during exposure to osmotic stress indicating that post-transcriptional regulation plays an important role in the adaptation of S. cerevisiae to osmotic stress.

摘要

酿酒酵母包含一个由17个己糖转运蛋白(HXT)基因组成的家族;只有9个已确定功能,其中一些功能仍未明确界定。尽管人们为表征己糖转运蛋白付出了巨大努力,但HXT6和HXT8 - 17的表达仍然是个谜。在自然环境中,酿酒酵母面临极端营养条件,包括超过40%(w/v)的糖、营养物质的耗尽以及温度和pH值的极端情况。利用HXT启动子 - lacZ融合技术,我们确定了HXT17基因表达的新条件;在pH 7.7而非pH 4.7的含有棉子糖和半乳糖的培养基中,HXT17启动子活性上调。我们证明,在存在非发酵碳源的情况下,HXT5、HXT13以及程度较轻的HXT15均被诱导表达。HXT1编码一种低亲和力转运蛋白,在短期渗透压休克实验中,当细胞暴露于含有40%葡萄糖的培养基时,HXT1启动子活性降低。然而,我们发现HXT1 mRNA转录本在渗透压应激条件下是稳定的。此外,HXT1 mRNA的稳定似乎并非基因特异性的,因为转录停止30分钟后,渗透压应激的酵母细胞中的mRNA比未应激的酵母细胞中的mRNA多四倍。因此,很大一部分酿酒酵母mRNA分子在暴露于渗透压应激期间的周转速率可能会降低,这表明转录后调控在酿酒酵母适应渗透压应激中起重要作用。

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