Stambuk B U, Panek A D, Crowe J H, Crowe L M, de Araujo P S
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, SP, Brazil.
Biochim Biophys Acta. 1998 Jan 8;1379(1):118-28. doi: 10.1016/s0304-4165(97)00087-1.
The expression of the high-affinity trehalose-H+ symport was investigated in various Saccharomyces cerevisiae strains and culture conditions. Previous kinetic studies of trehalose transport in yeast have revealed the existence of at least two different uptake mechanisms: a high-affinity trehalose-H+ symport activity repressed by glucose, and a constitutive low-affinity transport activity, a putative facilitated diffusion process. Exogenously added trehalose was not an inducer of the high-affinity transport activity, and a correlation between trehalose and maltose uptake by yeast cells was found. Our results indicate that the maltose-H+ symporters encoded by MAL11, MAL21, and MAL41 are not responsible for the trehalose transport activity. The analysis of both trehalose and maltose transport activities in wild-type and in laboratory strains with defined MAL genes showed that the trehalose-H+ symporter was under control of MAL regulatory genes. Our results also suggest that the recently characterized AGT1 gene of S. cerevisiae may encode the high-affinity trehalose-H+ symporter. During diauxic growth on glucose the transport activity was low during the first exponential phase of growth, increased as glucose was exhausted from the medium, and decreased again as the cells reached the late stationary phase. This pattern was coincident with that of the intracellular levels of trehalose. The strong correlation between these two parameters may be of physiological significance during adaptation of yeast cells to stress conditions.
在各种酿酒酵母菌株和培养条件下研究了高亲和力海藻糖-H⁺同向转运体的表达。先前对酵母中海藻糖转运的动力学研究表明,至少存在两种不同的摄取机制:一种受葡萄糖抑制的高亲和力海藻糖-H⁺同向转运活性,以及一种组成型低亲和力转运活性,推测为易化扩散过程。外源添加的海藻糖不是高亲和力转运活性的诱导剂,并且发现酵母细胞摄取海藻糖和麦芽糖之间存在相关性。我们的结果表明,由MAL11、MAL21和MAL41编码的麦芽糖-H⁺同向转运体与海藻糖转运活性无关。对野生型和具有确定MAL基因的实验室菌株中的海藻糖和麦芽糖转运活性进行分析表明,海藻糖-H⁺同向转运体受MAL调控基因的控制。我们的结果还表明,酿酒酵母最近鉴定的AGT1基因可能编码高亲和力海藻糖-H⁺同向转运体。在葡萄糖上进行双相生长期间,转运活性在生长的第一个指数期较低,随着培养基中葡萄糖耗尽而增加,并且随着细胞进入稳定后期而再次降低。这种模式与海藻糖的细胞内水平一致。在酵母细胞适应应激条件期间,这两个参数之间的强相关性可能具有生理意义。