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水稻钾离子通道β亚基的分子克隆与表达特性分析

Molecular cloning and expression characterization of a rice K+ channel beta subunit.

作者信息

Fang Z, Kamasani U, Berkowitz G A

机构信息

Plant Science Department, College of Agriculture and Natural Resources, University of Connecticut, Storrs 06269-4067, USA.

出版信息

Plant Mol Biol. 1998 Jul;37(4):597-606. doi: 10.1023/a:1005913629485.

Abstract

K+ channel proteins native to animal membranes have been shown to be composed of two different types of polypeptides: the pore-forming alpha subunit and the beta subunit which may be involved in either modulation of conductance through the channel, or stabilization and surface expression of the channel complex. Several cDNAs encoding animal K+ channel beta subunits have been recently cloned and sequenced. We report the molecular cloning of a rice plant homolog of these animal beta subunits. The rice cDNA (KOB1) described in this report encodes a 36 kDa polypeptide which shares 45% sequence identity with these animal K+ channel beta subunits. and 72% identity with the only other cloned plant (Arabidopsis thaliana) K+ channel beta subunit (KAB1). The KOB1 translation product was demonstrated to form a tight physical association with a plant K+ channel alpha subunit. These results are consistent with the conclusion that the KOB1 cDNA encodes a K+ channel beta subunit. Expression studies indicated that KOB1 protein is more abundant in leaves than in either reproductive structures or roots. Later-developing leaves on a rice plant were found to contain increasing levels of the protein with the flag leaf having the highest titer of KOB1. Leaf sheaths are known to accumulate excess K+ and act as reserve sources of this cation when new growth requires remobilization of K+. Leaf sheaths were found to contain higher levels of KOB1 protein than the blade portions of leaves. It was further determined that when K+ was lost from older leaves of plants grown on K+-deficient fertilizer, the loss of cellular K+ was associated with a decline in both KOB1 mRNA and protein. This finding represents the first demonstration (in either plants or animals) that changes in cellular K+ status may specifically alter expression of a gene encoding a K+ channel subunit.

摘要

动物细胞膜上的钾离子通道蛋白已被证明由两种不同类型的多肽组成

形成孔道的α亚基和β亚基,β亚基可能参与通过通道的电导调节,或通道复合物的稳定及表面表达。最近已克隆并测序了几个编码动物钾离子通道β亚基的cDNA。我们报道了这些动物β亚基的水稻同源物的分子克隆。本报告中描述的水稻cDNA(KOB1)编码一种36 kDa的多肽,与这些动物钾离子通道β亚基具有45%的序列同一性,与另一个已克隆的植物(拟南芥)钾离子通道β亚基(KAB1)具有72%的同一性。已证明KOB1翻译产物与植物钾离子通道α亚基形成紧密的物理结合。这些结果与KOB1 cDNA编码钾离子通道β亚基的结论一致。表达研究表明,KOB1蛋白在叶片中比在生殖结构或根中更丰富。发现水稻植株上较晚发育的叶片中该蛋白水平不断增加,旗叶中KOB1的含量最高。已知叶鞘会积累过量的钾离子,并在新生长需要钾离子重新 mobilization时作为这种阳离子的储备来源。发现叶鞘中KOB1蛋白的含量高于叶片的叶片部分。进一步确定,当在缺钾肥料上生长的植物的老叶中钾离子流失时,细胞钾离子的流失与KOB1 mRNA和蛋白的下降相关。这一发现首次证明(在植物或动物中)细胞钾离子状态的变化可能特异性地改变编码钾离子通道亚基的基因的表达。

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