State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, PR China.
J Exp Bot. 2013 Apr;64(7):1795-804. doi: 10.1093/jxb/ert039. Epub 2013 Feb 13.
Aluminium (Al)-activated citrate secretion plays an important role in Al resistance in a number of plant species, such as rice bean (Vigna umbellata). This study further characterized the regulation of VuMATE1, an aluminium-activated citrate transporter. Al stress induced VuMATE1 expression, followed by the secretion of citrate. Citrate secretion was specific to Al stress, whereas VuMATE1 expression was not, which could be explained by a combined regulation of VuMATE1 expression and Al-specific activation of VuMATE1 protein. Pre-treatment with a protein translation inhibitor suppressed VuMATE1 expression, indicating that de novo biosynthesis of proteins is required for gene expression. Furthermore, post-treatment with a protein translation inhibitor inhibited citrate secretion, indicating that post-transcriptional regulation of VuMATE1 is critical for citrate secretion. Protein kinase and phosphatase inhibitor studies showed that reversible phosphorylation was important not only for transcriptional regulation of VuMATE1 expression but also for post-translational regulation of VuMATE1 protein activity. These results suggest that citrate secretion is dependent on both transcriptional and post-transcriptional regulation of VuMATE1. Additionally, VuMATE1 promoter-β-glucuronidase fusion lines revealed that VuMATE1 expression was restricted to the root apex and was entirely Al induced, indicating the presence of cis-acting elements regulating root tip-specific and Al-inducible gene expression, which will be an important resource for genetic improvement of plant Al resistance.
铝(Al)激活的柠檬酸分泌在许多植物物种(如豇豆)的耐铝性中发挥着重要作用。本研究进一步表征了铝激活的柠檬酸转运蛋白 VuMATE1 的调控。铝胁迫诱导 VuMATE1 表达,随后柠檬酸分泌。柠檬酸分泌是特异于铝胁迫的,而 VuMATE1 表达不是,这可以通过 VuMATE1 表达的综合调控和铝特异性激活 VuMATE1 蛋白来解释。蛋白质翻译抑制剂预处理抑制 VuMATE1 表达,表明新合成的蛋白质是基因表达所必需的。此外,蛋白质翻译抑制剂的后处理抑制了柠檬酸的分泌,表明 VuMATE1 的转录后调控对于柠檬酸的分泌至关重要。蛋白激酶和磷酸酶抑制剂研究表明,可逆磷酸化不仅对 VuMATE1 表达的转录调控很重要,而且对 VuMATE1 蛋白活性的转录后调控也很重要。这些结果表明柠檬酸的分泌依赖于 VuMATE1 的转录和转录后调控。此外,VuMATE1 启动子-β-葡萄糖醛酸酶融合系揭示 VuMATE1 表达仅限于根尖,并且完全受铝诱导,表明存在调节根尖特异性和铝诱导基因表达的顺式作用元件,这将是遗传改良植物耐铝性的重要资源。