The interactome of soybean GmWRKY53 using yeast 2-hybrid library screening to saturation

Soybean GmWRKY53 functions in both biotic and abiotic stress signaling. Using GmWRKY53 as a bait yeast 2-hybrid library screening to saturation isolated multiple independent fragments for many interacting proteins, enabling delineation of minimal interacting domains and computation of a confidence score. Multiple independent clones coding for the LATE ELONGATED HYPOCOTYL clock protein GmLCL2 (MYB114) were isolated and the binding site for GmWRKY53 was mapped to 90 amino acids separate from the MYB domain. This suggests a direct input from the clock on GmWRKY53 activity. The GmWRKY53-interacting proteins also included 3 water stress-inducible AP2/ERF transcription factors. One of these (Glyma03g26310) is one of the most strongly water stress induced genes in soybean roots, suggesting that GmWRKY53/ERF complexes regulate water stress responses.

大豆GmWRKY53同时参与生物胁迫与非生物胁迫信号转导通路。以GmWRKY53为诱饵进行饱和酵母双杂交（yeast 2-hybrid）文库筛选，分离得到多个与多种互作蛋白对应的独立片段，可用于界定最小互作结构域并计算置信度评分。研究分离得到多个编码下胚轴延长延迟（LATE ELONGATED HYPOCOTYL）节律蛋白GmLCL2（MYB114）的独立克隆，并确定GmWRKY53的结合位点位于与MYB结构域相分离的90个氨基酸区域内。这表明生物钟可直接调控GmWRKY53的活性。GmWRKY53的互作蛋白还包含3种干旱胁迫诱导型AP2/ERF转录因子（AP2/ERF）。其中一个基因（Glyma03g26310）是大豆根系中受干旱胁迫诱导程度最高的基因之一，这提示GmWRKY53/ERF复合物可调控植物的干旱胁迫应答过程。