The uncommon function and mechanism of the common enzyme glyceraldehyde-3-phosphate dehydrogenase in the metamorphosis of Helicoverpa armigera.
The uncommon function and mechanism of the common enzyme glyceraldehyde-3-phosphate dehydrogenase in the metamorphosis of Helicoverpa armigera.
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棉铃虫变态过程中常见酶甘油醛-3-磷酸脱氢酶的异常功能及其机制。
DOI：
10.3389/fbioe.2022.1042867
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发表时间：
2022
影响因子：
5.7
通讯作者：
An, Shiheng
中科院分区：
文献类型：
--
作者：
Zhao, Wenli;Zhang, Bo;Geng, Zichen;Chang, Yanpeng;Wei, Jizhen;An, Shiheng
甘油醛-3-磷酸脱氢酶（GAPDH）是糖酵解的关键酶，常用作人类、小鼠和昆虫的内参基因。然而，GAPDH在昆虫发育，特别是变态过程中的作用尚未见报道。本研究以棉铃虫和草地贪夜蛾卵巢细胞系（Sf 9细胞）为材料，研究GAPDH在幼虫变态过程中的功能及其分子机制。结果表明，HaGAPDH与S. frugiperda和斜纹夜蛾Spodoptera litura。HaGAPDH在6龄幼虫中的转录高峰期分别为6L-3（表皮和中肠）和6L-1（脂肪体）d，qRT-PCR结果显示20 E和烯虫酯显著上调HaGAPDH的转录。HaGAPDH-GFP-His在Sf 9细胞中特异性定位于线粒体。在六龄幼虫中通过RNA干扰（RNAi）敲低HaGAPDH导致体重减轻、死亡率增加以及化蛹率和羽化率降低。HaGAPDH直接结合到可溶性海藻糖酶（HaTreh 1）物理和酵母双杂交，共免疫沉淀和共定位实验中的20 E处理。此外，HaGAPDH的敲低增加了Treh 1的活性，这反过来又降低了海藻糖含量，但增加了葡萄糖含量的幼虫。因此，这些数据表明GAPDH通过直接与HaTreh 1结合，将葡萄糖含量控制在正常范围内，以确保葡萄糖代谢和变态。
Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a key enzyme in glycolysis, is commonly used as an internal reference gene in humans, mice, and insects. However, the function of GAPDH in insect development, especially in metamorphosis, has not been reported. In the present study, Helicoverpa armigera and Spodoptera frugiperda ovarian cell lines (Sf9 cells) were used as materials to study the function and molecular mechanism of GAPDH in larval metamorphosis. The results showed that HaGAPDH was more closely related to GAPDH of S. frugiperda and Spodoptera litura. The transcript peaks of HaGAPDH in sixth instar larvae were 6L-3 (epidermal and midgut) and 6L-1 (fat body) days, and 20E and methoprene significantly upregulated the transcripts of HaGAPDH of larvae in qRT-PCR. HaGAPDH–GFP–His was specifically localized in mitochondria in Sf9 cells. Knockdown of HaGAPDH by RNA interference (RNAi) in sixth instar larvae resulted in weight loss, increased mortality, and decreases in the pupation rate and emergence rates. HaGAPDH is directly bound to soluble trehalase (HaTreh1) physically and under 20E treatment in yeast two-hybrid, coimmunoprecipitation, and colocalization experiments. In addition, knockdown of HaGAPDH increased the Treh1 activity, which in turn decreased the trehalose content but increased the glucose content in larvae. Therefore, these data demonstrated that GAPDH controlled the glucose content within the normal range to ensure glucose metabolism and metamorphosis by directly binding with HaTreh1.
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影响因子：
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作者：
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DOI：
10.1016/s0006-291x(03)01222-1
发表时间：
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影响因子：
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影响因子：
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作者：
Huang, Jinsha;Hao, Lijun;Wang, Tao
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影响因子：
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作者：
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