Vacuolar protein sorting 4 is required for silkworm metamorphosis

Vacuolar protein sorting 4 (Vps4) not only functions with its positive regulator vacuolar protein sorting 20-associated 1 (Vta1) in the multivesicular body (MVB) pathway but also participates alone in MVB-unrelated cellular processes. However, its physiological roles at the organism level remain rarely explored. We previously identified their respective homologues Bombyx mori Vps4 (BmVps4) and BmVta1 from the silkworm, a model organism for insect research. In this study, we performed fluorescence quantitative real-time PCR and Western blot to globally characterize the transcription and protein expression profiles of BmVps4 and BmVta1 during silkworm development and in different silkworm tissues and organs. The results showed that they were significantly up-regulated in metamorphosis, adulthood and embryogenesis relative to larval stages, and displayed a roughly similar tissue-and-organ specificity for transcriptions in silkworm larvae. Importantly, BmVps4 was down-regulated during the early period of the fifth instar, reaching the lowest level of transcription on Day 6, then up-regulated from Day 7 to the wandering, spinning and pupal stages, and down-regulated again in adulthood. Moreover, knocking down BmVps4 by RNA interference significantly inhibited silk gland growth, shortened spinning time, prolonged pupation, reduced pupal size and weight, and increased moth wing defects. Together, our data demonstrate the critical and broad requirements for BmVps4 in silkworm metamorphosis.