Major abiotic stress factors such as drought, salinity, hypoxia, and extreme temperatures along with rapid global climate change have had a huge negative impact on agricultural productivity. Understanding the abiotic stress-responsive molecular mechanisms and its associated proteins is extremely important to advance our knowledge towards developing multiple abiotic stress tolerance in plants. Firstly, basic understanding at transient level would be a vital foundation to accomplish this goal. Therefore, our present study aimed at understanding the sub-cellular localization of Eukaryotic Initiation Factor 4A-III (AteIF4A-III), a key DEAD-box RNA helicase, and Always Early 4 (AtALY4), an mRNA export factor, and their in vivo protein-protein interaction with major abiotic stress–associated proteins under control and multiple abiotic stress conditions. AteIF4A-III and AtALY4 were localized to the nucleus as evident by transient protoplast assay. AteIF4A-III has shown strong interaction with a negative regulator of multiple abiotic stresses, Stress Response Suppressor 1 (AtSTRS1) in Bi-FC assay. Further, the flow cytometry analysis has shown the strong interaction between them. Interestingly, under multiple abiotic stress treatment, the interacting partners were rapidly re-localized from nucleus to cytoplasm and cytoplasmic space. Similar results were observed when N- and C-terminal fusions of AteIF4A-III and AtALY4 treated under multiple abiotic stresses. Our study reveals that AteIF4A-III, AtALY4, and abiotic stress–associated protein AtSTRS1 are among the key proteins associated with multiple abiotic stress responses in plants.

中文翻译：

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拟南芥中多种非生物胁迫响应性 AteIF4A-III 和 AtALY4 蛋白的瞬时亚细胞定位和体内蛋白-蛋白相互作用研究

干旱、盐分、缺氧和极端温度等主要非生物胁迫因素以及全球气候的快速变化对农业生产力产生了巨大的负面影响。了解非生物胁迫响应分子机制及其相关蛋白质对于提高我们的知识以在植物中开发多种非生物胁迫耐受性极为重要。首先，对瞬态水平的基本理解将是实现这一目标的重要基础。因此，我们目前的研究旨在了解真核起始因子 4A-III (AteIF4A-III)（一种关键的死盒 RNA 解旋酶）和 Always Early 4（AtALY4）（一种 mRNA 输出因子）的亚细胞定位及其在在控制和多种非生物胁迫条件下与主要非生物胁迫相关蛋白的体内蛋白质-蛋白质相互作用。AteIF4A-III 和 AtALY4 定位于细胞核，如瞬时原生质体测定所示。AteIF4A-III 在 Bi-FC 测定中显示出与多种非生物胁迫的负调节因子 Stress Response Suppressor 1 (AtSTRS1) 的强相互作用。此外，流式细胞术分析表明它们之间存在很强的相互作用。有趣的是，在多重非生物胁迫处理下，相互作用的伙伴迅速从细胞核重新定位到细胞质和细胞质空间。当 AteIF4A-III 和 AtALY4 的 N 端和 C 端融合体在多种非生物胁迫下处理时，观察到类似的结果。我们的研究表明，AteIF4A-III、AtALY4 和非生物胁迫相关蛋白 AtSTRS1 是与植物中多种非生物胁迫反应相关的关键蛋白。