A TFIIIA-type zinc finger protein confers multiple abiotic stress tolerances in transgenic rice (Oryza sativa L.)
A TFIIIA-type zinc finger protein confers multiple abiotic stress tolerances in transgenic rice (Oryza sativa L.)
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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
DOI：
10.1007/s11103-012-9955-5
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发表时间：
2012-10-01
影响因子：
5.1
通讯作者：
Zhang, Hongsheng
中科院分区：
文献类型：
--
作者：
Huang, Ji;Sun, Shujing;Zhang, Hongsheng
TFIIIA型锌指转录因子参与植物发育和非生物胁迫反应。大多数TFIIIA型锌指蛋白是转录抑制因子，因为它们的氨基酸序列中存在一个耳朵基序。在这项工作中，我们发现ZFP182，一种TFIIIA型锌指蛋白，在细胞核中形成同源二聚体，并在酵母细胞中表现出反式激活活性。缺失分析表明，反式激活需要C末端的富亮氨酸区域。过表达ZFP 182显著提高了转基因水稻的耐盐性、耐冷性和抗旱性等多种非生物胁迫的耐性。ZFP 182的过表达促进了转基因水稻中亲和性渗透调节物质的积累，如游离脯氨酸和可溶性糖。在胁迫条件下，ZFP 182激活了编码吡咯烷-5-羧酸合成酶的OsP5CS和OsLEA3的表达，而在正常和胁迫条件下，OsDREB1A和OsDREB1B都受到ZFP 182的调控。有趣的是，定点突变实验表明ZFP 182启动子中的DRE类似元件参与了脱水诱导ZFP 182的表达。酵母双杂交实验表明，ZFP 182与多种核糖体蛋白相互作用，包括ZIURP1，一种融合了核糖体蛋白L40的泛素。通过双分子荧光互补和His下拉实验进一步证实了ZFP 182和ZIURP1在体内和体外的相互作用。我们的研究为理解TFIIIA型锌指转录因子介导抗逆性的机制提供了新的线索，并为提高作物的抗逆性提供了候选基因。
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.