Quintero F J, Blatt M R
Laboratory of Plant Physiology and Biophysics, University of London, Wye College, UK.
FEBS Lett. 1997 Sep 29;415(2):206-11. doi: 10.1016/s0014-5793(97)01125-3.
Transport of K+ in higher plants, as in bacteria and fungi, is mediated by two broad classes of transport proteins that operate in the millimolar and micromolar K+ concentration ranges. A search of the Expressed Sequence Tag database using amino acid consensus sequences for the K+ transporters HAK1 from Schwanniomyces and Kup of Escherichia coli yielded two homologous sequences for Arabidopsis. Cloning and sequencing of these genes gave single open reading frames for the putative transporters, AtKT1 and AtKT2, with predicted molecular weights of 79 and 88 kDa. The predicted gene products showed a high degree of homology at the amino acid level (56% identity) and exhibited significant hydrophobic stretches in their N-terminal halves, consistent with 12 membrane-spanning, alpha-helical domains. Database searches using AtKT1 and AtKT2 identified 10 additional sequences in Arabidopsis as well as additional homologous sequences in the plant species Oryza and Allium, the bacterium Lactococcus lactis, and in Homo sapiens. Expression of AtKT2 rescued growth on low millimolar [K+] in Saccharomyces cerevisiae carrying deletions for the genes encoding the K+ transporters TRK1 and TRK2. Rescue was associated with a 2-fold stimulation of Rb+ uptake and was sensitive to competition with external Na+ but not to extracellular pH, indicating that the gene encodes a low-affinity K+ transporter. These and additional results suggest that AtKT1 and AtKT2 belong to a superfamily of cation transporters that have been conserved through evolution.
与细菌和真菌一样,高等植物中钾离子的转运由两大类转运蛋白介导,它们分别在毫摩尔和微摩尔钾离子浓度范围内发挥作用。利用来自施万酵母属的钾离子转运蛋白HAK1和大肠杆菌的Kup的氨基酸共有序列搜索表达序列标签数据库,得到了拟南芥的两个同源序列。对这些基因进行克隆和测序,得到了推定转运蛋白AtKT1和AtKT2的单一开放阅读框,预测分子量分别为79 kDa和88 kDa。预测的基因产物在氨基酸水平上具有高度同源性(同一性为56%),并且在其N端半部表现出明显的疏水延伸,这与12个跨膜α螺旋结构域一致。使用AtKT1和AtKT2进行数据库搜索,在拟南芥中鉴定出另外10个序列,以及在植物物种水稻和葱、细菌乳酸乳球菌和人类中的其他同源序列。AtKT2的表达挽救了携带钾离子转运蛋白TRK1和TRK2编码基因缺失的酿酒酵母在低毫摩尔[K+]条件下的生长。挽救与铷离子摄取增加2倍有关,并且对外部钠离子的竞争敏感,但对细胞外pH不敏感,这表明该基因编码一种低亲和力钾离子转运蛋白。这些以及其他结果表明,AtKT1和AtKT2属于一个在进化过程中保守的阳离子转运蛋白超家族。