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一种 TFIIIA 型锌指蛋白赋予转基因水稻(Oryza sativa L.)多种非生物胁迫耐受性。

A TFIIIA-type zinc finger protein confers multiple abiotic stress tolerances in transgenic rice (Oryza sativa L.).

机构信息

State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Agriculture, Nanjing Agricultural University, Nanjing 210095, China.

出版信息

Plant Mol Biol. 2012 Oct;80(3):337-50. doi: 10.1007/s11103-012-9955-5. Epub 2012 Aug 29.

Abstract

The TFIIIA-type zinc finger transcription factors are involved in plant development and abiotic stress responses. Most TFIIIA-type zinc finger proteins are transcription repressors due to existence of an EAR-motif in their amino acid sequences. In this work, we found that ZFP182, a TFIIIA-type zinc finger protein, forms a homodimer in the nucleus and exhibits trans-activation activity in yeast cells. The deletion analysis indicated that a Leu-rich region at C-terminus is required for the trans-activation. Overexpression of ZFP182 significantly enhanced multiple abiotic stress tolerances, including salt, cold and drought tolerances in transgenic rice. Overexpression of ZFP182 promotes accumulation of compatible osmolytes, such as free proline and soluble sugars, in transgenic rice. ZFP182 activates the expression of OsP5CS encoding pyrroline-5-carboxylate synthetase and OsLEA3 under stress conditions, while OsDREB1A and OsDREB1B were regulated by ZFP182 under both normal and stress conditions. Interestingly, site-directed mutagenesis assay showed that DRE-like elements in ZFP182 promoter are involved in dehydration-induced expression of ZFP182. The yeast two-hybrid assay revealed that ZFP182 interacted with several ribosomal proteins including ZIURP1, an ubiquitin fused to ribosomal protein L40. The in vivo and in vitro interactions of ZFP182 and ZIURP1 were further confirmed by bimolecular fluorescence complementation and His pull-down assays. Our studies provide new clues in the understanding of the mechanisms for TFIIIA-type zinc finger transcription factor mediated stress tolerance and a candidate gene for improving stress tolerance in crops.

摘要

TFIIIA 型锌指转录因子参与植物发育和非生物胁迫响应。由于其氨基酸序列中存在 EAR 基序,大多数 TFIIIA 型锌指蛋白是转录抑制剂。在这项工作中,我们发现 TFIIIA 型锌指蛋白 ZFP182 在核内形成同源二聚体,并在酵母细胞中表现出转录激活活性。缺失分析表明 C 末端富含亮氨酸的区域对于转录激活是必需的。ZFP182 的过表达显著增强了多种非生物胁迫耐受性,包括转基因水稻的耐盐、耐寒和耐旱性。ZFP182 的过表达促进了转基因水稻中相容渗透物(如游离脯氨酸和可溶性糖)的积累。ZFP182 在胁迫条件下激活编码吡咯啉-5-羧酸合成酶的 OsP5CS 和 OsLEA3 的表达,而 OsDREB1A 和 OsDREB1B 则受 ZFP182 在正常和胁迫条件下的调控。有趣的是,定点突变实验表明,ZFP182 启动子中的 DRE 样元件参与了 ZFP182 的脱水诱导表达。酵母双杂交实验表明,ZFP182 与几种核糖体蛋白相互作用,包括 ZIURP1,一种与核糖体蛋白 L40 融合的泛素。ZFP182 和 ZIURP1 的体内和体外相互作用进一步通过双分子荧光互补和 His 下拉实验得到证实。我们的研究为理解 TFIIIA 型锌指转录因子介导的胁迫耐受性机制提供了新的线索,并为提高作物的胁迫耐受性提供了一个候选基因。

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