Characterization of the CsPNG1 gene from cucumber and its function in response to salinity stress
Characterization of the CsPNG1 gene from cucumber and its function in response to salinity stress
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黄瓜 CsPNG1 基因的特征及其响应盐胁迫的功能
DOI：
10.1016/j.plaphy.2020.02.027
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发表时间：
2020-05-01
影响因子：
6.5
通讯作者：
Guo, Shi-Rong
中科院分区：
文献类型：
--
作者：
Hou, Kun;Wang, Yu;Guo, Shi-Rong
肽：N-聚糖酶（PNGase; EC 3.5.1.52）是一种去糖基化酶，负责内质网中错误折叠的糖蛋白的去糖基化。然而，PNGase在植物中的作用在很大程度上是未知的。本研究克隆了黄瓜N-聚糖酶（CsPNG 1）基因，并对其功能进行了初步研究。CsPNG 1基因编码的氨基酸含有典型的转氨酶（TGase）催化三联体结构域，属于“TGase超家族”。亚细胞定位结果表明，CsPNG 1定位于细胞膜和细胞核。启动子序列分析和qPCR检测结果表明，CsPNG 1能够响应多种非生物胁迫和激素处理。酵母单杂交实验揭示了转录因子CsGT-3b和CsPNG 1启动子之间的相互作用。重要的是，CsPNG 1在烟草中的过表达提高了转基因植物对盐胁迫的耐受性。CsPNG 1与CsRAD 23家族蛋白相互作用，CsRAD 23蛋白C端乌巴结构域与CsPNG 1结合，提示CsPNG 1参与了ER相关降解途径（ERAD）。综上所述，我们的研究表明CsPNG 1基因在提高植物耐盐性方面发挥了积极作用，这些结果为进一步分析CsPNG 1基因在非生物胁迫和ERAD中的功能奠定了基础。
Peptide: N-glycanase (PNGase; EC 3.5.1.52) is a deglycosylation enzyme that is responsible for deglycosylating misfolded glycoproteins in the endoplasmic reticulum. However, the role of PNGase in plants is largely unknown. Here, we cloned and characterized the function of peptide: N-glycanase (CsPNG1) from cucumber. The amino acid encoded by CsPNG1 gene contained a typical transglutaminase (TGase) catalytic triad domain and belonged to the "TGase superfamily". Subcellular localization showed that CsPNG1 was located in the cell membrane and nucleus. Promoter sequence analysis and qPCR tests showed that CsPNG1 could respond to a variety of abiotic stresses and hormone treatments. Yeast one-hybrid assays revealed the interaction between the transcription factor CsGT-3b and CsPNG1 promoter. Importantly, overexpression of CsPNG1 in tobacco increased the tolerance to salt stress of transgenic plants. In addition, CsPNG1 interacted with CsRAD23 family proteins and the C-terminal UBA domain of CsRAD23 protein was responsible for binding to CsPNG1, indicating that CsPNG1 was involved in the ER-associated degradation pathway (ERAD). Taken together, our study demonstrated that CsPNG1 plays a positive role in improving plant salt tolerance, and these findings might provide a basis for further functional analysis of CsPNG1 genes in abiotic stress and ERAD.