N-Terminal asparagine amidohydrolase is a component of the ubiquitin-dependent N-end rule pathway of protein degradation that has been implicated in a variety of physiological functions, including the sensing of heme, oxygen, nitric oxide, selective elimination of misfolded proteins and the repair of DNA. We identified the Apis cerana cerana N-terminal asparagine amidohydrolase (AccNtan1) gene from A. cerana cerana and investigated its role in oxidation resistance. Multiple sequence alignments and phylogenetic analysis revealed that N-terminal asparagine amidohydrolase is highly conserved in insect species. Quantitative real-time polymerase chain reaction analysis indicated that the expression levels of AccNtan1 were significantly lower in the wing, honey sac and abdomen than in other tissues and were significantly higher in early stages of development, including the larva, prepupa and pink-eyed pupa stages, than in later stages. We further observed that AccNtan1 expression was induced by several environmental stressors, including aberrant temperature, H₂O₂, UV, heavy metals and pesticides. Moreover, a bacteriostatic assay suggested that overexpression of AccNtan1 enhances the resistance of bacteria to oxidative stress. In addition, knockdown of AccNtan1 using RNA interference significantly affected the expression levels of most antioxidant genes and the activity levels of several antioxidant enzymes. Thus, we hypothesize that AccNtan1 plays important roles in environmental stress responses and antioxidative processes.

中文翻译：

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中华蜜蜂N-末端天冬酰胺酰胺水解酶（AccNtan1）在氧化应激中的作用。

N端天冬酰胺酰胺水解酶是蛋白质降解的泛素依赖性N端规则途径的一个组成部分，已与多种生理功能有关，包括血红素，氧气，一氧化氮的感应，错折叠蛋白的选择性消除以及DNA修复。我们确定了中华蜜蜂N-末端天冬酰胺酰胺水解酶（AccNtan1）基因从A.中华蜜蜂和耐氧化性研究其作用。多个序列比对和系统发育分析表明，N-末端天冬酰胺酰胺水解酶在昆虫物种中高度保守。实时定量聚合酶链反应分析表明，AccNtan1的表达水平在机翼，蜜囊和腹部的显着低于其他组织，并且在发育的早期阶段（包括幼虫，前pu和粉红色眼的stages期）显着高于后期。我们进一步观察到，AccNtan1表达是由多种环境胁迫诱导的，包括温度异常，H ₂ O ₂，紫外线，重金属和农药。此外，抑菌实验表明，AccNtan1的过表达增强了细菌对氧化应激的抵抗力。此外，AccNtan1的敲低使用RNA干扰显着影响大多数抗氧化剂基因的表达水平和几种抗氧化酶的活性水平。因此，我们假设AccNtan1在环境应激反应和抗氧化过程中起重要作用。