Schachtman D P, Kumar R, Schroeder J I, Marsh E L
Department of Botany, University of Adelaide, Adelaide, SA 5005 Australia.
Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):11079-84. doi: 10.1073/pnas.94.20.11079.
The transport of cations across membranes in higher plants plays an essential role in many physiological processes including mineral nutrition, cell expansion, and the transduction of environmental signals. In higher plants the coordinated expression of transport mechanisms is essential for specialized cellular processes and for adaptation to variable environmental conditions. To understand the molecular basis of cation transport in plant roots, a Triticum aestivum cDNA library was used to complement a yeast mutant deficient in potassium (K+) uptake. Two genes were cloned that complemented the mutant: HKT1 and a novel cDNA described in this report encoding a cation transporter, LCT1 (low-affinity cation transporter). Analysis of the secondary structure of LCT1 suggests that the protein contains 8-10 transmembrane helices and a hydrophilic amino terminus containing sequences enriched in Pro, Ser, Thr, and Glu (PEST). The transporter activity was assayed using radioactive isotopes in yeast cells expressing the cDNA. LCT1 mediated low-affinity uptake of the cations Rb+ and Na+, and possibly allowed Ca2+ but not Zn2+ uptake. LCT1 is expressed in low abundance in wheat roots and leaves. The precise functional role of this cation transporter is not known, although the competitive inhibition of cation uptake by Ca2+ has parallels to whole plant and molecular studies that have shown the important role of Ca2+ in reducing Na+ uptake and ameliorating Na+ toxicity. The structure of this higher plant ion transport protein is unique and contains PEST sequences.
在高等植物中,阳离子跨膜运输在许多生理过程中起着至关重要的作用,包括矿质营养、细胞扩张以及环境信号转导。在高等植物中,运输机制的协调表达对于特殊的细胞过程和适应多变的环境条件至关重要。为了了解植物根系中阳离子运输的分子基础,利用普通小麦的cDNA文库对钾(K⁺)吸收缺陷的酵母突变体进行互补。克隆到了两个能够互补该突变体的基因:HKT1和本报告中描述的一个编码阳离子转运蛋白LCT1(低亲和性阳离子转运蛋白)的新cDNA。对LCT1二级结构的分析表明,该蛋白含有8 - 10个跨膜螺旋以及一个富含脯氨酸(Pro)、丝氨酸(Ser)、苏氨酸(Thr)和谷氨酸(Glu)(PEST)的亲水性氨基末端。在表达该cDNA的酵母细胞中使用放射性同位素测定转运蛋白活性。LCT1介导阳离子Rb⁺和Na⁺的低亲和性吸收,可能也允许Ca²⁺吸收,但不允许Zn²⁺吸收。LCT1在小麦根和叶中的表达量较低。尽管Ca²⁺对阳离子吸收的竞争性抑制与全株和分子研究有相似之处,这些研究表明Ca²⁺在减少Na⁺吸收和减轻Na⁺毒性方面具有重要作用,但这种阳离子转运蛋白的确切功能作用尚不清楚。这种高等植物离子转运蛋白的结构独特,并且含有PEST序列。