State Key Laboratory of Genetic Engineering, Institute of Plant Biology, School of Life Sciences, Fudan University, Shanghai, 200433, China.
Plant J. 2013 Oct;76(2):274-86. doi: 10.1111/tpj.12287. Epub 2013 Aug 12.
The unfolded protein response (UPR) plays important roles in plant development and plant-pathogen interactions, as well as in plant adaptation to adverse environmental stresses. Previously bZIP28 and bZIP60 have been identified as important UPR regulators for mitigating the endoplasmic reticulum (ER) stress in Arabidopsis thaliana. Here we report the biological function of NAC103 in a novel transcriptional regulatory cascade, connecting bZIP60 to the UPR downstream genes in Arabidopsis. Expression of NAC103 was induced by ER stress, and was completely abolished in the bZIP60 null mutant. A new ER stress-responsive cis-element UPRE-III (TCATCG) on the NAC103 promoter was identified, and trans-activation of UPRE-III by bZIP60 was confirmed in both yeast cells and Arabidopsis protoplasts. The direct binding of bZIP60 to UPRE-III-containing DNA was also demonstrated in an electrophoretic mobility shift assay. NAC103 formed homodimers in yeast two-hybrid and bimolecular fluorescence complementation assays. It had transcriptional activation activity and was localized in the nucleus. Over-expression of NAC103 had pleiotropic effects on plant growth, and induced expression of several UPR downstream genes in Arabidopsis under normal growth conditions. The activation of UPR gene promoters by NAC103 was also confirmed in effector/reporter protoplast assays. Thus, our study demonstrates a transcriptional regulatory cascade in which NAC103 relays ER stress signals from bZIP60 to UPR downstream genes through a newly identified ER stress cis-element (UPRE-III) and transcriptional activation activity of its encoded protein NAC103.
未折叠蛋白反应 (UPR) 在植物发育和植物-病原体相互作用中以及在植物适应不利环境压力方面发挥着重要作用。以前已经鉴定出 bZIP28 和 bZIP60 是缓解拟南芥内质网 (ER) 应激的重要 UPR 调节剂。在这里,我们报告了 NAC103 在一个新的转录调控级联中的生物学功能,该级联将 bZIP60 与拟南芥 UPR 下游基因连接起来。NAC103 的表达受 ER 应激诱导,并且在 bZIP60 缺失突变体中完全被消除。在 NAC103 启动子上鉴定到一个新的 ER 应激反应顺式元件 UPRE-III (TCATCG),并在酵母细胞和拟南芥原生质体中证实了 bZIP60 对 UPRE-III 的转录激活。在电泳迁移率变动分析中还证实了 bZIP60 与含有 UPRE-III 的 DNA 的直接结合。NAC103 在酵母双杂交和双分子荧光互补测定中形成同源二聚体。它具有转录激活活性并且位于细胞核中。在正常生长条件下,NAC103 的过表达对植物生长具有多效性,并诱导拟南芥中几个 UPR 下游基因的表达。在效应子/报告子原生质体测定中也证实了 NAC103 对 UPR 基因启动子的激活。因此,我们的研究表明,在转录调控级联中,NAC103 通过新鉴定的 ER 应激顺式元件 (UPRE-III) 和其编码蛋白 NAC103 的转录激活活性,将 ER 应激信号从 bZIP60 传递到 UPR 下游基因。