Bombyx mori lipase-1 reduces viral replication and proliferation by inhibiting the expression of the DNA-binding protein (dbp) gene.
Chen Haiyu H, Ding Xiangrui X, Wang Jinyang J, Zhao Qiaoling Q et al.
Bombyx mori lipase-1 (Bmlipase-1) has strong anti-BmNPV activity, but the mechanism by which Bmlipase-1 resists BmNPV invasion has not been elucidated so far. In this paper, based on the construction of recombinant viruses overexpressing Bmlipase-1, we analyzed the effects of the recombinant viruses on the genes related to recombinant virus replication as well as viral replication and proliferation after the recombinant viruses infected BmN cells. In the presence of the virus, Bmlipase-1 localizes to the cell nucleus. Quantitative polymerase chain reaction (qPCR), northern blotting and western blot analyses showed that Bmlipase-1 inhibited the expression of BmNPV DNA-binding protein (dbp) as well as the replication and proliferation of the recombinant virus. DBP is a key factor for normal viral DNA replication and for the stabilization of nascent viral DNA. A biotin-labeled probe was synthesized around the c-Myb transcription factor binding site in the upstream promoter region of dbp and analyzed by EMSA with prokaryotically expressed and purified Bmlipase-1. It was found that Bmlipase-1 could bind to CACTTCAATT in the dbp promoter region, while the activity of the dbp promoter lacking CACTTCAATT was significantly reduced. In conclusion, Bmlipase-1 can bind to the CACTTCAATT motif on the dbp promoter in vitro, and this motif contributes positively to the activity of the dbp promoter. Overexpression of Bmlipase-1 significantly downregulates dbp expression and inhibits viral replication. Based on these findings, we propose the following hypothesis: Bmlipase-1 may inhibit viral replication by binding to the CACTTCAATT motif and interfering with the transcriptional regulation of dbp. However, this regulatory model requires further validation in the context of viral infection. This paper provides a new theoretical basis for the prevention and control of lepidopteran insect viruses, and also opens up a new way to study the antiviral mechanism of the silkworm, Bombyx mori.