Cao Y, Ward J M, Kelly W B, Ichida A M, Gaber R F, Anderson J A, Uozumi N, Schroeder J I, Crawford N M
Department of Biology, University of California, San Diego, La Jolla 92093-0116, USA.
Plant Physiol. 1995 Nov;109(3):1093-106. doi: 10.1104/pp.109.3.1093.
K+ channels play diverse roles in mediating K+ transport and in modulating the membrane potential in higher plant cells during growth and development. Some of the diversity in K+ channel functions may arise from the regulated expression of multiple genes encoding different K+ channel polypeptides. Here we report the isolation of a novel Arabidopsis thaliana cDNA (AKT2) that is highly homologous to the two previously identified K+ channel genes, KAT1 and AKT1. This cDNA mapped to the center of chromosome 4 by restriction fragment length polymorphism analysis and was highly expressed in leaves, whereas AKT1 was mainly expressed in roots. In addition, we show that diversity in K+ channel function may be attributable to differences in expression levels. Increasing KAT1 expression in Xenopus oocytes by polyadenylation of the KAT1 mRNA increased the current amplitude and led to higher levels of KAT1 protein, as assayed in western blots. The increase in KAT1 expression in oocytes produced shifts in the threshold potential for activation to more positive membrane potentials and decreased half-activation times. These results suggest that different levels of expression and tissue-specific expression of different K+ channel isoforms can contribute to the functional diversity of plant K+ channels. The identification of a highly expressed, leaf-specific K+ channel homolog in plants should allow further molecular characterization of K+ channel functions for physiological K+ transport processes in leaves.
钾离子通道在高等植物细胞生长和发育过程中介导钾离子运输以及调节膜电位方面发挥着多种作用。钾离子通道功能的一些多样性可能源于编码不同钾离子通道多肽的多个基因的调控表达。在此,我们报告了拟南芥一种新的cDNA(AKT2)的分离,它与之前鉴定的两个钾离子通道基因KAT1和AKT1高度同源。通过限制性片段长度多态性分析,该cDNA定位于第4号染色体的中心,且在叶片中高表达,而AKT1主要在根中表达。此外,我们表明钾离子通道功能的多样性可能归因于表达水平的差异。通过对KAT1 mRNA进行聚腺苷酸化处理来增加非洲爪蟾卵母细胞中KAT1的表达,可增加电流幅度,并导致更高水平的KAT1蛋白,这在蛋白质免疫印迹分析中得到证实。卵母细胞中KAT1表达的增加使激活阈值电位向更正的膜电位偏移,并缩短了半激活时间。这些结果表明,不同钾离子通道亚型的不同表达水平和组织特异性表达可导致植物钾离子通道的功能多样性。鉴定出植物中一种高表达、叶特异性的钾离子通道同源物,应有助于进一步从分子层面表征叶片中钾离子通道在生理钾离子运输过程中的功能。