Vacuolar ATPase subunit H is essential for the survival and moulting of Locusta migratoria manilensis
Vacuolar ATPase subunit H is essential for the survival and moulting of Locusta migratoria manilensis
复制标题
液泡 ATP 酶 H 亚基对于马尼拉飞蝗的生存和蜕皮至关重要
DOI：
10.1111/j.1365-2583.2012.01147.x
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发表时间：
2012-08-01
影响因子：
2.6
通讯作者：
Xia, Y.
中科院分区：
文献类型：
--
作者：
Li, C.;Xia, Y.
液泡(H+)-ATPase(V-ATPase)作为一个生电泵，将质子从细胞质输送到细胞外液中，产生细胞负膜电压。Vhasfd编码的V-ATPase亚基H是V-ATPase活性所必需的。本研究克隆了东亚飞虱V-ATPase亚基H基因，命名为Lm-Vhasfd。全长为2018个碱基，开放阅读框编码515个氨基酸残基。半定量逆转录聚合酶链式反应(RT-PCR)显示，LM-Vhasfd在血淋巴、中肠、躯干和腿部组织中的转录水平较高，而在脂肪体和头部组织中的转录水平相对较低。注射特定的双链RNA(DsRNA)后，Lm-Vhasfd mRNA、V-ATPase活性和ATP浓度均显著降低。生物测定表明，沉默LM-Vhasfd会导致个体死亡和各种蜕皮缺陷。RNAi突变体在注射后11d的累积死亡率为96.7%，显著高于野生型蝗虫。这些RNAi表型表明，Lm-Vhasfd对东亚飞虱的生长和蜕皮是必需的。
Vacuolar (H+)-ATPase (V-ATPase) functions as an electrogenic pump, transporting protons from the cytoplasm to the extracellular fluid to generate cell-negative membrane voltage. The V-ATPase subunit H, encoded by Vhasfd, is required for V-ATPase activity. In this study, the gene encoding V-ATPase subunit H from Locusta migratoria manilensis was cloned, and designated as Lm-Vhasfd. The complete cDNA sequence is 2018 bp, with an open reading frame encoding 515 amino acid residues. Semi-quantitative reverse transcription PCR (RT-PCR) showed that Lm-Vhasfd transcription is high in the haemolymph, midgut, trunk and leg, but relatively low in the fat body and head tissues. Injection with a specific double-strand RNA (dsRNA) led to a significant decrease in Lm-Vhasfd mRNA, V-ATPase enzyme activity and ATP concentration. Bioassays showed that silencing Lm-Vhasfd led to the death of individuals and various moulting defects. The accumulative mortality of the RNA interference (RNAi) mutant 11 days post-injection was 96.7%, which was conspicuously higher than that seen in wild type locusts. These RNAi phenotypes demonstrate that Lm-Vhasfd is essential for the growth and moulting of L. migratoria manilensis.