Vacuolar ATPase subunit H is essential for the survival and moulting of Locusta migratoria manilensis.
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.
Genetic Engineering Research Center, School of Bioengineering, Chongqing Engineering Research Center for Fungal Insecticide, The Key Laboratory of Gene Function and Expression Regulation, Chongqing University, Chongqing, China.
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D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D048188:Locusta migratoria / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D018978:Molting / Q000235:genetics*; D009928:Organ Specificity; D010802:Phylogeny; D034622:RNA Interference; D017422:Sequence Analysis, DNA / Q000379:methods; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics / Q000378:metabolism / Q000502:physiology
D025262:Vacuolar Proton-Translocating ATPases