E93 promotes transcription of RHG genes to initiate apoptosis during Drosophila salivary gland metamorphosis.

20-hydroxyecdysone (20E) induced transcription factor E93 is important for larval-adult transition, which functions in programmed cell death of larval obsolete tissues, and the formation of adult new tissues. However, the apoptosis-related genes directly regulated by E93 are still ambiguous. In this study, an E93 mutation fly strain was obtained by clustered regularly interspaced palindromic repeats (CRISPR) / CRISPR-associated protein 9-mediated long exon deletion to investigate whether and how E93 induces apoptosis during larval tissues metamorphosis. The transcriptional profile of E93 was consistent with 3 RHG (rpr, hid, and grim) genes and the effector caspase gene drice, and all their expressions peaked at the initiation of apoptosis during the degradation of salivary glands. The transcription expression of 3 RHG genes decreased and apoptosis was blocked in E93 mutation salivary gland during metamorphosis. In contrast, E93 overexpression promoted the transcription of 3 RHG genes, and induced advanced apoptosis in the salivary gland. Moreover, E93 not only enhance the promoter activities of the 3 RHG genes in Drosophila Kc cells in vitro, but also in the salivary gland in vivo. Our results demonstrated that 20E induced E93 promotes the transcription of RHG genes to trigger apoptosis during obsolete tissues degradation at metamorphosis in Drosophila.