Identification of Host Molecules Involved in the Proliferation of Nucleopolyhedrovirus in Bombyx mori

Abstract

The Bombyx mori nucleopolyhedrovirus (BmNPV), a foodborne infectious virus, is the pathogen causing nuclear polyhedrosis and high lethality in the silkworm. In this study, we characterized the molecules involved in BmNPV–silkworm interaction by RNA sequencing of the fat body isolated from the virus-susceptible strain P50. Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation showed that the upregulated differentially expressed genes (DEGs) were mainly involved in translation, signal transduction, folding, sorting, and degradation, as well as transport and catabolism, while the downregulated DEGs were predominantly enriched in the metabolism of carbohydrates, amino acids, and lipids at 72 h post BmNPV infection. Knockout of the upregulated somatomedin-B and thrombospondin type-1 domain-containing protein, probable allantoicase, trifunctional purine biosynthetic protein adenosine-3, and Psl and pyoverdine operon regulator inhibited the proliferation of BmNPV, while knockout of the downregulated clip domain serine protease 3 and carboxylesterase clade H, member 1 promoted it. The molecules herein identified provide a foundation for developing strategies and designing drugs against BmNPV.

Cytoskeleton Protein BmACT1 Is Potential for the Autophagic Function and Nuclear Localization of BmAtg4b in Bombyx mori

“…However, the roles of BmAtg4a and BmAtg4b in viral proliferation suffer from a lack of investigation. After overexpression of BmAtg4a for 48 h, the BmN cells were infected with BmNPV (MOI = 5) for a further 48 h [ 25 ]. Monitored by gene copies of BmNPV ie1 , gp64 , and helicase, as well as the variation of the green fluorescence of recombinant BmNPV-EGFP, overexpression of BmAtg4a significantly reduced the viral replication ( Figure 3 A,B,B’).…”

“…A full-length sequence of Egfp was cloned into a pFastBac Dual Expression Vector, and then the plasmid was transformed into DH10Bac competent cells integrated with the BmNPV genome [ 24 ]. Subsequently, the bacmid DNA was extracted and transfected into BmN cells to generate the P1 (progeny 1) virus, and then the P2 virus was obtained by infecting BmN cells with the P1 virus for 5 days; the TCID 50 of a titer of P2 BmNPV was determined [ 25 ].…”

“…BmN cells co-transfected with empty pB-CRISPR and A3-Helper vectors were used as control. After transfection for 48 h, 600 μg/mL zeocin (Gibco, Waltham, MA, USA, R25001) was added to the cells, which were chemically selected and maintained for 1 month to remove the cells unsuccessfully integrated with the CRISPR/Cas9 system in the genome to obtain a stable cell line [ 25 ].…”

“…Western blotting was performed as previously described [ 25 ]. Primary antibodies were used to detect the protein levels of EGFP (TransGen, Beijing, China, HT801; 1:4000), His (Cell Signaling Technology, USA, 12698s; 1:3000), His (TransGen, Beijing, HT501; 1:4000), HA (Santa Cruz Biotechnology, Santa Cruz, CA, USA, sc-7392; 1:3000), FLAG (Sigma, USA, F1804; 1:3000), V5 (Sigma, Saint Louis, MO, USA, H1029; 1:3000), and BmAtg8 (ABclonal Biotechnology, Shanghai, China; 1:3000).…”

Bombyx mori UFL1 facilitates BmNPV proliferation by regulating host cell apoptosis through PERK

“…Among these viruses, BmNPV is the most prevalent threat to silkworm and sericulture (Lü et al, 2018). Therefore, BmNPV‐silkworm interaction has been a hot field for both basic research and agriculture in recent decades (Xu et al, 2022).…”