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鲁氏接合酵母Trk1是一种高效的钾离子转运蛋白,可赋予酵母细胞高锂耐受性。

Zygosaccharomyces rouxii Trk1 is an efficient potassium transporter providing yeast cells with high lithium tolerance.

作者信息

Zimmermannova Olga, Salazar Ana, Sychrova Hana, Ramos Jose

机构信息

Department of Membrane Transport, Institute of Physiology Czech Academy of Sciences, CZ-14220 Prague, Czech Republic.

Departament of Microbiology, E.T.S.I.A.M., University of Cordoba, E-14071 Cordoba, Spain.

出版信息

FEMS Yeast Res. 2015 Jun;15(4):fov029. doi: 10.1093/femsyr/fov029. Epub 2015 May 27.

Abstract

Zygosaccharomyces rouxii is an osmotolerant yeast growing in the presence of high concentrations of salts and/or sugars. The maintenance of intracellular potassium homeostasis is essential for osmostress adaptation. Zygosaccharomyces rouxii is endowed with only one typical potassium transporter (ZrTrk1). We characterized ZrTrk1 activity and its contribution to various physiological parameters in detail. Our results show that ZrTrk1 is a high-affinity K(+) transporting system efficiently discriminating between K(+) and Li(+) and indicate the presence of another, currently unknown K(+) importing system with a low affinity in Z. rouxii cells. Upon ZrTrk1 heterologous expression in Saccharomyces cerevisiae, it confers cells with a remarkably high lithium tolerance (even to wild-type strains) due to preventing Li(+) influx into cells, and is able to complement a plasma-membrane hyperpolarization and cell sensitivity to cationic compounds caused by the lack of endogenous K(+) transporters. Intracellular pH measurements with pHluorin, whose coding sequence was integrated into the genome, showed that the expression of ZrTrk1 also complements a decrease in intracellular pH in S. cerevisiae trk1Δ trk2Δ cells. Our data corroborate a tight connection between potassium and proton transporters in yeasts and provide new insights into Z. rouxii cation homeostasis and the basis of its high osmotolerance.

摘要

鲁氏接合酵母是一种耐渗透压酵母,能在高浓度盐和/或糖存在的情况下生长。维持细胞内钾离子稳态对于适应渗透压胁迫至关重要。鲁氏接合酵母仅具有一种典型的钾离子转运蛋白(ZrTrk1)。我们详细表征了ZrTrk1的活性及其对各种生理参数的贡献。我们的结果表明,ZrTrk1是一种高效区分钾离子和锂离子的高亲和力钾离子转运系统,并表明在鲁氏接合酵母细胞中存在另一种目前未知的低亲和力钾离子导入系统。在酿酒酵母中异源表达ZrTrk1后,由于阻止锂离子流入细胞,它赋予细胞极高的锂耐受性(甚至对野生型菌株也是如此),并且能够弥补由于缺乏内源性钾离子转运蛋白而导致的质膜超极化和细胞对阳离子化合物的敏感性。用编码序列整合到基因组中的pHluorin进行细胞内pH测量表明,ZrTrk1的表达还能弥补酿酒酵母trk1Δ trk2Δ细胞内pH的降低。我们的数据证实了酵母中钾离子和质子转运蛋白之间的紧密联系,并为鲁氏接合酵母的阳离子稳态及其高渗透压耐受性的基础提供了新的见解。

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