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来自耐旱禾本科植物斯氏鼠尾粟的一种脱水响应小GTP结合蛋白(Rab2)的特性分析

Characterization of a desiccation-responsive small GTP-binding protein (Rab2) from the desiccation-tolerant grass Sporobolus stapfianus.

作者信息

O'Mahony P J, Oliver M J

机构信息

Plant Stress and Water Conservation Laboratory, USDA ARS, Lubbock, TX 79401, USA.

出版信息

Plant Mol Biol. 1999 Mar;39(4):809-21. doi: 10.1023/a:1006183431854.

Abstract

We have used differential display to detect altering mRNA levels in response to desiccation and rehydration in leaves of the desiccation tolerant grass Sporobolus stapfianus. One of the RT-PCR products identified was used to isolate a cDNA of 999 bp which encodes a protein of 210 amino acids (predicted size 23 kDa). This protein displays considerable sequence similarity to mammalian and plant Rab2, a small GTP-binding protein, possessing several conserved motifs common to these regulatory proteins. Sporobolus Rab2 was expressed in Escherichia coli yielding a protein with an apparent molecular mass of ca. 30 kDa which was shown to have the ability to bind GTP. Rab2 transcript accumulated early in response to a decrease in relative water content (RWC) and remained high even in dried leaves. Rehydration of desiccated leaves resulted in a decrease in levels within 3 h of rewetting, with a brief increase at ca. 12 h. Accumulation of Rab2 transcript was also evident during drying and rehydration of the roots of S. stapfianus, as well as in leaves of the desiccation-sensitive grass Sporobolus pyramidalis. Earlier work on S. stapfianus concluded that the plant hormone ABA has little effect on inducing desiccation tolerance, however Rab2 transcript does exhibit a small increase in accumulation in response to exogenous ABA. A possible role for Rab2 with respect to desiccation tolerance and damage repair is discussed.

摘要

我们利用差异显示技术检测了耐旱禾本科植物斯氏鼠尾粟叶片在脱水和复水过程中mRNA水平的变化。所鉴定出的一个逆转录聚合酶链反应(RT-PCR)产物被用于分离一个999 bp的互补DNA(cDNA),该cDNA编码一个由210个氨基酸组成的蛋白质(预测大小为23 kDa)。这种蛋白质与哺乳动物和植物的Rab2(一种小GTP结合蛋白)具有相当高的序列相似性,拥有这些调节蛋白共有的几个保守基序。斯氏鼠尾粟Rab2在大肠杆菌中表达,产生一种表观分子量约为30 kDa的蛋白质,该蛋白质被证明具有结合GTP的能力。Rab2转录本在相对含水量(RWC)降低时早期积累,即使在干燥叶片中仍保持高水平。脱水叶片复水导致复水3小时内水平下降,在约12小时时有短暂升高。Rab2转录本在斯氏鼠尾粟根的干燥和复水过程中以及在对脱水敏感的禾本科植物圆锥鼠尾粟的叶片中积累也很明显。早期对斯氏鼠尾粟的研究得出结论,植物激素脱落酸(ABA)对诱导耐旱性影响不大,然而Rab2转录本在响应外源ABA时积累确实有小幅增加。本文讨论了Rab2在耐旱性和损伤修复方面可能的作用。

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