Multiple regulation of Arabidopsis AtGST11 gene expression by four transcription factors under abiotic stresses
Multiple regulation of Arabidopsis AtGST11 gene expression by four transcription factors under abiotic stresses
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DOI：
10.1111/j.1399-3054.2012.01699.x
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发表时间：
2013-05-01
影响因子：
6.4
通讯作者：
Ezaki, Bunichi
中科院分区：
文献类型：
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作者：
Kouno, Takafumi;Ezaki, Bunichi
拟南芥谷胱甘肽S-转移酶（GST; E.C. 2.5.1.18）基因AtGST 11受许多环境应激的调节，包括金属毒性、氧化应激和极端温度。为了阐明这一调控机制，我们利用酵母单杂交（Y1 H）技术分离并鉴定了两个转录因子[（TF），P1-1和P1-3]。这些TF分别编码推定的bZIP TF（AtbZIP 30）和乙烯反应元件结合因子2（AtERF 2）。Y1 H和电泳迁移率变动分析（EMSA）表明，两种蛋白均与AtGST 11基因的5-上游区域结合。启动子活性测定表明，这两个TF上调AtGST 11。此外，发现另外两个先前分离的TF（#13和#43）分别编码C3 HC 4型RING指（DAL 1）和HomeBox蛋白6（AtHB 6）下调。AtGST 11基因在野生型株系（Col-0）的表达增加镉，铜和铝，但降低冷，热和二酰胺处理。为了确定每个TF如何与AtGST 11基因的环境调控相关，使用来自两个破坏的突变株系（DAL 1和AtHB 6）或过表达株系（oxAtERF 2）的RNA样品在各种压力下进行定量实时聚合酶链反应。结果表明，DAL 1和AtHB 6作为阻遏物下调AtGST 11的表达，AtERF 2作为诱导物上调AtGST 11在热胁迫下的表达。
The Arabidopsis thaliana glutathione S-transferase (GST; E.C. 2.5.1.18) gene, AtGST11, is regulated by a number of environmental stresses, including metal toxicities, oxidative stress and extremes of temperature. To clarify the mechanisms of this regulation, two transcription factors [(TFs), P1-1 and P1-3] were isolated by yeast one-hybrid (Y1H) analysis and characterized. These TFs encoded a putative bZIP TF (AtbZIP30) and ethylene-response element-binding factor 2 (AtERF2), respectively. Y1H and electrophoresis mobility shift assay (EMSA) showed that both proteins bound to the 5-upstream region of the AtGST11 gene. Promoter activity assays indicated that both TFs upregulate AtGST11. In addition, another two previously isolated TFs (#13 and #43), encoding C3HC4-type RING finger (DAL1) and Homeobox protein 6 (AtHB6), respectively, were found to downregulate. Expression of the AtGST11 gene in a wild-type line (Col-0) was increased by Cd, Cu and Al, but decreased by cold, heat and diamide treatment. To determine how each TF relates to the environmental regulation of the AtGST11 gene, quantitative real-time polymerase chain reaction was performed using RNA samples derived from two disrupted mutant lines (DAL1 and AtHB6) or an over-expression line (oxAtERF2) under various stresses. The results indicated that DAL1 and AtHB6 downregulated the expression of AtGST11 under, most of the stresses tested as, repressors and AtERF2 upregulated the expression under heat stress as inducer.