Polidori Emanuela, Ceccaroli Paola, Saltarelli Roberta, Guescini Michele, Menotta Michele, Agostini Deborah, Palma Francesco, Stocchi Vilberto
Istituto di Ricerca sull'Attività Motoria, Università degli Studi di Urbino Carlo Bo, Via I Maggetti 26, 61029 Urbino, Italy.
Fungal Genet Biol. 2007 Mar;44(3):187-98. doi: 10.1016/j.fgb.2006.08.001. Epub 2006 Sep 26.
Here, we report the first evidence of a hexose transporter gene, Tbhxt1, in the ectomycorrhizal ascomycete Tuber borchii Vittadini. The protein encoded by Tbhxt1 functionally complements the hxt-null mutant Saccharomyces cerevisiae EBYVW.4000. TBHXT1 has a strong preference for d-glucose (K(m)=38+/-10 microM) over d-fructose (K(m)=16+/-5mM) and uncoupling experiments indicate that TBHXT1 catalyzes the transport via a proton-symport mechanism. The investigations on the substrate specificity reveal that TBHXT1 also imports d-mannose, and the use of deoxyglucose analogues shows that the hydroxyl groups at C1, C3 and C4 are important for substrate recognition. Tbhxt1 is not regulated by fructose, but it reaches its highest level of expression at 3mM glucose and is repressed by very high glucose concentration. Prolonged carbon starvation condition upregulates Tbhxt1, while its expression remains at basal level in the ectomycorrhizal tissue. The mode of regulation of Tbhxt1 is consistent with its role as a high-affinity d-glucose transporter.
在此,我们报告了外生菌根子囊菌波氏块菌(Tuber borchii Vittadini)中己糖转运蛋白基因Tbhxt1的首个证据。Tbhxt1编码的蛋白在功能上互补了hxt基因缺失的酿酒酵母(Saccharomyces cerevisiae)EBYVW.4000突变体。与d-果糖(K(m)=16±5mM)相比,TBHXT1对d-葡萄糖具有很强的偏好性(K(m)=38±10μM),解偶联实验表明TBHXT1通过质子同向转运机制催化转运。对底物特异性的研究表明,TBHXT1也能转运d-甘露糖,使用脱氧葡萄糖类似物表明C1、C3和C4位的羟基对于底物识别很重要。Tbhxt1不受果糖调控,但在3mM葡萄糖时达到最高表达水平,并受到非常高的葡萄糖浓度的抑制。长时间的碳饥饿条件会上调Tbhxt1,而其在菌根组织中的表达维持在基础水平。Tbhxt1的调控模式与其作为高亲和力d-葡萄糖转运蛋白的作用一致。