Maresova Lydie, Sychrova Hana
Department of Membrane Transport, Institute of Physiology, Academy of Sciences, CR, Videnska 1083, 142 20 Prague 4, Czech Republic.
Yeast. 2006 Dec;23(16):1167-71. doi: 10.1002/yea.1424.
AtChx17p is a putative K(+)/H(+) exchanger from Arabidopsis thaliana, expressed in the roots and probably involved in K(+) acquisition and homeostasis. AtCHX17 cDNA complements the phenotypes of the kha1Delta mutation in S. cerevisiae cells: a growth defect at increased pH and hygromycin sensitivity. The localization of GFP-tagged AtChx17 protein in yeast cells is similar to that of ScKha1p: a bold dotted pattern inside the cells resembling the Golgi fluorescence markers. These results show that (a) the proteins AtChx17 and ScKha1 could have similar functions and (b) S. cerevisiae kha1 deletion mutants could serve for the heterologous expression and characterization of plant transporters. The results of this work are evidence that a S. cerevisiae strain with deletions of genes encoding alkali-metal-cation/H(+) antiporters (i.e. Nha1p, Nhx1p, Kha1p) could be an ideal tool for expression and functional analysis of any type of similar plant antiporters (plasma membrane, endosomal/prevacuolar and Golgi).
AtChx17p是拟南芥中一种假定的钾离子/氢离子交换体,在根中表达,可能参与钾离子的获取和稳态维持。AtCHX17 cDNA能够弥补酿酒酵母细胞中kha1Delta突变的表型:在pH升高时生长缺陷以及对潮霉素敏感。绿色荧光蛋白标记的AtChx17蛋白在酵母细胞中的定位与ScKha1p相似:细胞内呈现粗大的点状模式,类似于高尔基体荧光标记。这些结果表明:(a)AtChx17和ScKha1蛋白可能具有相似功能;(b)酿酒酵母kha1缺失突变体可用于植物转运蛋白的异源表达和特性研究。这项工作的结果证明,缺失编码碱金属阳离子/氢离子反向转运蛋白(即Nha1p、Nhx1p、Kha1p)基因的酿酒酵母菌株可能是用于表达和功能分析任何类型相似植物反向转运蛋白(质膜、内体/液泡前体和高尔基体)的理想工具。