Wang Hongfei, Xie Yucai, Wang Xiaoqing, Geng Xiaoshan, Gao Lijun
College of Life Sciences, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian, Shandong, China.
College of Life Sciences, Shandong First Medical University & Shandong Academy of Medical Sciences, Taian, Shandong, China.
Comp Biochem Physiol B Biochem Mol Biol. 2023 Jan;263:110796. doi: 10.1016/j.cbpb.2022.110796. Epub 2022 Aug 13.
Receptors for Activated C Kinase 1 (RACK1s) are a kind of multifunction scaffold protein that plays an important role in cell signal transductions and animal development. However, the function of RACK1 in the Chinese honeybee Apis cerana cerana is little known. Here, we isolated and identified a RACK1 gene from Apis cerana cerana, named AccRACK1. By bioinformatic analysis, we revealed a high nucleic acid homology between AccRACK1 and RACK1 of Apis cerana. RT-qPCR analyses demonstrated AccRACK1 was mostly expressed in 3rd instar larvae, darked-eyed pupae and adults (one and thirty days post-emergence), suggesting it might participate in the development of A. cerana cerana. Moreover, the expression of AccRACK1 was highest in the thorax, followed by the venom gland. Compared to the blank control group, AccRACK1 was induced by 24 and 44 °C, HgCl and pesticides (paraquat, pyridaben and methomyl) but inhibited by 14 °C, HO, UV light and cyhalothrin. Additionally, 0.05, 0.1, 1, 5 and 10 mg/ml PPN (juvenile hormone analogue pyriproxyfen) could promote the expression of AccRACK1, with 1 mg/ml showing the highest upregulation, suggesting it was regulated by hormones. Further study found that after knockdown of AccRACK1 by RNAi, the expression of the eukaryotic initiation factor 6 of A. cerana cerana (AcceIF6), an initiation factor regulating the initiation of translation, was inhibited, indicating AccRACK1 might affect cellular responses by translation. These findings, taken together, suggest AccRACK1 is involved in the development and responses to abiotic stresses of A. cerana cerana, and therefore, it may be of critical importance to the survival of A. cerana cerana.
活化C激酶1受体（RACK1s）是一类多功能支架蛋白，在细胞信号转导和动物发育中发挥重要作用。然而，RACK1在中国蜜蜂中华蜜蜂中的功能鲜为人知。在此，我们从中华蜜蜂中分离并鉴定了一个RACK1基因，命名为AccRACK1。通过生物信息学分析，我们揭示了AccRACK1与意大利蜜蜂RACK1之间具有高度的核酸同源性。RT-qPCR分析表明，AccRACK1主要在3龄幼虫、黑眼蛹和成虫（羽化后1天和30天）中表达，表明它可能参与中华蜜蜂的发育。此外，AccRACK1在胸部的表达最高，其次是毒腺。与空白对照组相比，AccRACK1在24和44℃、HgCl以及农药（百草枯、哒螨灵和灭多威）处理下被诱导，但在14℃、H₂O₂、紫外线和氯氟氰菊酯处理下受到抑制。此外，0.05、0.1、1、5和10mg/ml的PPN（保幼激素类似物吡丙醚）可促进AccRACK1的表达，其中1mg/ml的上调幅度最高，表明它受激素调节。进一步研究发现，通过RNAi敲低AccRACK1后，中华蜜蜂真核起始因子6（AcceIF6）的表达受到抑制，AcceIF6是一种调节翻译起始的起始因子，这表明AccRACK1可能通过翻译影响细胞反应。综上所述，这些发现表明AccRACK1参与中华蜜蜂的发育以及对非生物胁迫的反应，因此，它可能对中华蜜蜂的生存至关重要。
