Key message

WRKY50 from A. thaliana requires WT-boxes at target gene promoters for activation and binding.

Abstract

Based on the genome-wide prediction of WRKY50 target genes and the similarity of a WRKY50 binding site to WT-boxes in microbe-associated molecular pattern (MAMP)-responsive cis-regulatory modules (CRM), four WT-box containing CRMs from the promoter region of three WRKY50 target genes were investigated for their interaction with WRKY50. These target genes are DJ1E, WRKY30 and ATBBE4. Two of the four CRMs, one from DJ1E and one from WRKY30, were able to activate reporter gene expression in the presence of WRKY50. Activation requires the WT-boxes GGACTTTT, GGACTTTG from DJ1E and GGACTTTC from WRKY30. WRKY50 does not activate a second CRM from WRKY30 and the CRM from ATBBE4, both containing the WT-box TGACTTTT. In vitro gel-shift assays demonstrate WT-box-specific binding of the WRKY50 DNA-binding domain to all four CRMs. This work shows a high flexibility of WRKY50 binding site recognition beyond the classic W-box TTGACC/T.

中文翻译：

---

拟南芥 WRKY50 转录因子在靶基因启动子处的异常 DNA 结合特性

关键信息

来自拟南芥的 WRKY50 需要靶基因启动子处的 WT-box 来激活和结合。

抽象的

基于对 WRKY50 靶基因的全基因组预测以及 WRKY50 结合位点与微生物相关分子模式 (MAMP) 响应的顺式调节模块 (CRM) 中 WT-box 的相似性，四个含有来自 CRM 的 WT-box研究了三个 WRKY50 靶基因的启动子区域与 WRKY50 的相互作用。这些靶基因是DJ1E、WRKY30和ATBBE4。四种 CRM 中的两种，一种来自DJ1E，一种来自WRKY30，能够在 WRKY50 存在的情况下激活报告基因表达。激活需要来自 DJ1E 的 WT-box GGACTTTT、GGACTTTG和来自WRKY30 的GGACTTTC. WRKY50 不会激活来自 WRKY30 的第二个 CRM 和来自ATBBE4的CRM ，两者都包含 WT-box TGACTTTT。体外凝胶迁移试验证明了 WRKY50 DNA 结合域与所有四种 CRM 的 WT-box 特异性结合。这项工作显示了 WRKY50 结合位点识别的高度灵活性，超出了经典的 W-box TTGACC/T。