Abstract
Bombyx mori nucleopolyhedrovirus (BmNPV) is a primary pathogen of silkworm (B. mori) that causes severe economic losses each year. However, the molecular mechanisms of silkworm-BmNPV interactions, especially the silkworm proteins that can interact with the virus, are still largely unknown. In this study, the total and membrane proteins of silkworm midguts were displayed using one- and two-dimensional electrophoresis. A virus overlay assay was used to detect B. mori proteins that specifically bind to BmNPV particles. Twelve proteins were located and identified using mass spectrometry, and the different expression of the corresponding genes in BmNPV susceptible and resistant silkworm strains also indicated their involvement in BmNPV infection. The 12 proteins are grouped based on their potential roles in viral infection, for example, endocytosis, intracellular transportation, and host responses. Based on these results, we hypothesize the following: I) vacuolar ATP synthase catalytic subunit A and subunit B may be implicated in the process of the membrane fusion of virus and the release of the nucleocapsid into cytoplasm; II) actin, enolase and phosphoglycerate kinase are cytoskeleton associated proteins and may play an important role in BmNPV intracellular transportation; III) mitochondrial prohibitin complex protein 2, ganglioside-induced differentiation-associated protein, calreticulin, regucalcin-like isoform X1 and 60 kDa heat shock protein are involved in cell apoptosis regulation during BmNPV infection in larvae midguts; IV) ribosomal P0 may be associated with BmNPV infection by regulating gene expression of BmNPV; V) arginine kinase has a role in the antiviral activities against BmNPV. Our work should prove informative by providing multiple protein targets and a novel direction to investigate the molecular mechanisms of the interactions between silkworms and BmNPV.
Highlights
The silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), is an economically important insect for production of silk and recombinant proteins, and a good model of the Lepidoptera [1]
budded virus (BV) infect a broad range of cell types and transmit virus among insect tissues within an infected larva, whereas occlusion-derived virus (ODV) are contained in polyhedrons and form occlusion bodies (OBs) which infect only columnar epithelial cells of the insect midguts and are required for the oral transmission of virus between insect hosts [4, 5]
It has been reported that B. mori serine protease-2, lipase-1 and alkaline trypsin protein purified from the digestive juice of B. mori larvae showed strong antiviral activity to Bombyx mori nucleopolyhedrovirus (BmNPV) in vitro [6,7,8]
Summary
The silkworm, Bombyx mori L. (Lepidoptera: Bombycidae), is an economically important insect for production of silk and recombinant proteins, and a good model of the Lepidoptera [1]. Virus overlay assays were performed in the screening for BmNPV binding proteins from larval midguts of B. mori.
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