Leandro Maria José, Spencer-Martins Isabel, Gonçalves Paula
Centro de Recursos Microbiológicos (CREM), Department of Life Sciences, Faculty of Sciences and Technology, New University of Lisbon, 2829-516 Caparica, Portugal.
Microbiology (Reading). 2008 Jun;154(Pt 6):1646-1655. doi: 10.1099/mic.0.2007/015511-0.
Two glucose/xylose transporter genes from Candida intermedia were recently cloned and characterized: GXF1, which encodes a glucose/xylose facilitator; and GXS1, which encodes a glucose/xylose proton symporter. Here we report the functional expression of these transporters in Saccharomyces cerevisiae. While Gxf1p seems to be fully functional in S. cerevisiae, the symporter Gxs1p exhibits very low glucose/xylose transport activity, which could not be ascribed to insufficient production of the protein or incorrect subcellular localization. In addition, co-expression of glucose/xylose facilitators with Gxs1p strongly reduced GXS1 mRNA levels, and consequently symport activity, in glucose-grown, but not in ethanol-grown, cells. The observed decrease in GXS1 transcript levels seems to be related to an enhanced glucose influx mediated by glucose facilitator protein(s), and not to a specific interaction between Gxs1p and other transporters. We found GXS1 mRNA levels to be severely reduced as a result of glucose addition, and we show that this effect takes place at the level of GXS1 mRNA stability. Our results suggest that a decrease in mRNAs encoding high-affinity/active sugar transport systems may be a widespread and conserved mechanism in yeasts, limiting expression of these proteins whenever their activity is dispensable.
最近从中间假丝酵母中克隆并鉴定了两个葡萄糖/木糖转运蛋白基因:GXF1,其编码一种葡萄糖/木糖易化子;以及GXS1,其编码一种葡萄糖/木糖质子同向转运体。在此我们报道了这些转运蛋白在酿酒酵母中的功能表达。虽然Gxf1p在酿酒酵母中似乎具有完全功能,但同向转运体Gxs1p表现出非常低的葡萄糖/木糖转运活性,这不能归因于蛋白质产量不足或亚细胞定位错误。此外,葡萄糖/木糖易化子与Gxs1p共表达在葡萄糖培养而非乙醇培养的细胞中强烈降低了GXS1 mRNA水平,进而降低了同向转运活性。观察到的GXS1转录本水平下降似乎与葡萄糖易化蛋白介导的葡萄糖内流增强有关,而不是与Gxs1p和其他转运蛋白之间的特异性相互作用有关。我们发现添加葡萄糖会导致GXS1 mRNA水平严重降低,并且我们表明这种效应发生在GXS1 mRNA稳定性水平。我们的结果表明,编码高亲和力/活性糖转运系统的mRNA减少可能是酵母中一种广泛存在且保守的机制,在这些蛋白的活性可有可无时限制其表达。