Nuclear localization of the dehydrin OpsDHN1 is determined by histidine-rich motif.
Nuclear localization of the dehydrin OpsDHN1 is determined by histidine-rich motif.
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DOI：
10.3389/fpls.2015.00702
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发表时间：
2015
影响因子：
5.6
通讯作者：
Jiménez-Bremont JF
中科院分区：
文献类型：
--
作者：
Hernández-Sánchez IE;Maruri-López I;Ferrando A;Carbonell J;Graether SP;Jiménez-Bremont JF
仙人掌OpsDHN 1蛋白属于一个大家族的无序和高度亲水性的蛋白质，称为晚期胚胎发生丰富（莱亚）蛋白，其积累在胚胎发生的后期阶段和响应非生物胁迫。在此，我们提出了在体内OpsDHN 1的亚细胞定位的N-末端GFP翻译融合，我们的结果揭示了细胞质和核定位的GFP：：OpsDHN 1蛋白在烟草本氏表皮细胞。此外，使用双分子荧光互补（BiFC）方法证明了OpsDHN 1在植物中的二聚体组装。为了理解富含组氨酸的基序的体内作用，产生OpsDHN 1-ΔHis形式，并分别通过GFP融合和BiFC测定来测定其亚细胞定位和二聚体能力。我们发现，OpsDHN 1富含组氨酸的基序的缺失限制了其定位于细胞质，但不影响二聚体的形成。此外，OpsDHN 1蛋白中S片段的缺失影响其核定位。我们的数据表明，富含组氨酸的基序和S-节段的缺失表现出类似的效果，阻止OpsDHN 1进入细胞核。基于这些结果，提出了富含组氨酸的基序作为OpsDHN 1核定位的靶向元件。
The cactus OpsDHN1 dehydrin belongs to a large family of disordered and highly hydrophilic proteins known as Late Embryogenesis Abundant (LEA) proteins, which accumulate during the late stages of embryogenesis and in response to abiotic stresses. Herein, we present the in vivo OpsDHN1 subcellular localization by N-terminal GFP translational fusion; our results revealed a cytoplasmic and nuclear localization of the GFP::OpsDHN1 protein in Nicotiana benthamiana epidermal cells. In addition, dimer assembly of OpsDHN1 in planta using a Bimolecular Fluorescence Complementation (BiFC) approach was demonstrated. In order to understand the in vivo role of the histidine-rich motif, the OpsDHN1-ΔHis version was produced and assayed for its subcellular localization and dimer capability by GFP fusion and BiFC assays, respectively. We found that deletion of the OpsDHN1 histidine-rich motif restricted its localization to cytoplasm, but did not affect dimer formation. In addition, the deletion of the S-segment in the OpsDHN1 protein affected its nuclear localization. Our data suggest that the deletion of histidine-rich motif and S-segment show similar effects, preventing OpsDHN1 from getting into the nucleus. Based on these results, the histidine-rich motif is proposed as a targeting element for OpsDHN1 nuclear localization.