Ferreira Célia, van Voorst Frank, Martins António, Neves Luisa, Oliveira Rui, Kielland-Brandt Morten C, Lucas Cândida, Brandt Anders
Carlsberg Laboratory, Copenhagen Valby, Denmark.
Mol Biol Cell. 2005 Apr;16(4):2068-76. doi: 10.1091/mbc.e04-10-0884. Epub 2005 Feb 9.
Glycerol and other polyols are used as osmoprotectants by many organisms. Several yeasts and other fungi can take up glycerol by proton symport. To identify genes involved in active glycerol uptake in Saccharomyces cerevisiae we screened a deletion mutant collection comprising 321 genes encoding proteins with 6 or more predicted transmembrane domains for impaired growth on glycerol medium. Deletion of STL1, which encodes a member of the sugar transporter family, eliminates active glycerol transport. Stl1p is present in the plasma membrane in S. cerevisiae during conditions where glycerol symport is functional. Both the Stl1 protein and the active glycerol transport are subject to glucose-induced inactivation, following identical patterns. Furthermore, the Stl1 protein and the glycerol symporter activity are strongly but transiently induced when cells are subjected to osmotic shock. STL1 was heterologously expressed in Schizosaccharomyces pombe, a yeast that does not contain its own active glycerol transport system. In S. pombe, STL1 conferred the ability to take up glycerol against a concentration gradient in a proton motive force-dependent manner. We conclude that the glycerol proton symporter in S. cerevisiae is encoded by STL1.
甘油和其他多元醇被许多生物体用作渗透保护剂。几种酵母和其他真菌可以通过质子同向转运吸收甘油。为了鉴定酿酒酵母中参与主动甘油摄取的基因,我们筛选了一个缺失突变体文库,该文库包含321个编码具有6个或更多预测跨膜结构域的蛋白质的基因,以检测其在甘油培养基上生长受损的情况。编码糖转运蛋白家族成员的STL1基因的缺失消除了主动甘油转运。在甘油同向转运功能正常的条件下,Stl1p存在于酿酒酵母的质膜中。Stl1蛋白和主动甘油转运都遵循相同的模式,受到葡萄糖诱导的失活作用。此外,当细胞受到渗透冲击时Stl1蛋白和甘油同向转运蛋白活性会强烈但短暂地被诱导。STL1在粟酒裂殖酵母中进行了异源表达,粟酒裂殖酵母是一种自身不具备主动甘油转运系统的酵母。在粟酒裂殖酵母中,STL1赋予了其以质子动力依赖的方式逆浓度梯度吸收甘油的能力。我们得出结论,酿酒酵母中的甘油质子同向转运蛋白由STL1编码。