Zhang Yuanying, Liu Yaling, Guo Xulei, Li Yalu, Gao Hongru, Guo Xingqi, Xu Baohua
State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China; School of Basic Medical Sciences, Taishan Medical University, Taian, Shandong 271000, PR China.
School of Basic Medical Sciences, Taishan Medical University, Taian, Shandong 271000, PR China.
Insect Biochem Mol Biol. 2014 Oct;53:1-12. doi: 10.1016/j.ibmb.2014.06.007. Epub 2014 Jul 5.
Small heat shock proteins (sHSPs) play an important role in protecting against stress-induced cell damage and fundamental physiological processes. In this study, we identified an intronless sHsp gene from Apis cerana cerana (AccsHsp22.6). The open reading frame of AccsHsp22.6 was 585 bp and encoded a 194 amino acid protein. Furthermore, a 2064 bp 5'-flanking region was isolated, and potential transcription factor binding sites associated with development and stress response were identified. Quantitative PCR and western blot analyses demonstrated that AccsHsp22.6 was detected at higher levels in the midgut than in other tissues tested, and it is highly expressed during the shift to different development stages. Moreover, AccsHsp22.6 was significantly up-regulated by abiotic and biotic stresses, such as 4 °C, 16 °C, 42 °C, cyhalothrin, pyridaben, H2O2, UV, CdCl2, 20-hydroxyecdysone and Ascosphaera apis treatments. However, AccsHsp22.6 was slightly repressed by other stresses, including 25 °C, phoxim, paraquat and HgCl2 treatments. The recombinant AccsHSP22.6 also exhibited significant temperature tolerance, antioxidation and molecular chaperone activity. In addition, we found that knockdown of AccsHsp22.6 by RNA interference remarkably reduced temperature tolerance in A. cerana cerana. Taken together, these results suggest that AccsHsp22.6 plays an essential role in the development stages and defence against cellular stress.
小热休克蛋白（sHSPs）在抵御应激诱导的细胞损伤和基础生理过程中发挥着重要作用。在本研究中，我们从中华蜜蜂（Apis cerana cerana）中鉴定出一个无内含子的sHsp基因（AccsHsp22.6）。AccsHsp22.6的开放阅读框为585 bp，编码一个由194个氨基酸组成的蛋白质。此外，我们分离出了一个2064 bp的5'侧翼区域，并鉴定出了与发育和应激反应相关的潜在转录因子结合位点。定量PCR和蛋白质印迹分析表明，AccsHsp22.6在中肠中的检测水平高于其他测试组织，并且在向不同发育阶段转变时高表达。此外，AccsHsp22.6在非生物和生物胁迫下显著上调，如4℃、16℃、42℃、氯氟氰菊酯、哒螨灵、H2O2、紫外线、CdCl2、20-羟基蜕皮激素和蜜蜂球囊菌处理。然而，AccsHsp22.6在其他胁迫下略有抑制，包括25℃、辛硫磷、百草枯和HgCl2处理。重组AccsHSP22.6还表现出显著的温度耐受性、抗氧化和分子伴侣活性。此外，我们发现通过RNA干扰敲低AccsHsp22.6可显著降低中华蜜蜂的温度耐受性。综上所述，这些结果表明AccsHsp22.6在发育阶段和抵御细胞应激中起着至关重要的作用。
