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Identification and Functional Analysis of BmNPV-Interacting Proteins From Bombyx mori (Lepidoptera) Larval Midgut
Shang-Zhi Zhang1,2, Lin-Bao Zhu1,2, Dong Yu1,2
1School of Life Sciences, Anhui Agricultural University, Hefei, China.
Abstract:
Bombyx mori nucleopolyhedrovirus (BmNPV) is a major pathogen causing severe economic loss. However, the molecular mechanism of silkworm resistance to BmNPV and the interactions of this virus with the host during infection remain largely unclear. To explore the virus-binding proteins of silkworms, the midgut subcellular component proteins that may interact with BmNPV were analyzed in vitro based on one- and two-dimensional electrophoresis and far-western blotting combined with mass spectrometry (MS). A total of 24 proteins were determined to be specifically bound to budded viruses (BVs) in two subcellular fractions (mitochondria and microsomes). These proteins were involved in viral transportation, energy metabolism, apoptosis and viral propagation, and they responded to BmNPV infection with different expression profiles in different resistant strains. In particular, almost all the identified proteins were downregulated in the A35 strain following BmNPV infection. Interestingly, there were no virus-binding proteins identified in the cytosolic fraction of the silkworm midgut. Two candidate proteins, RACK1 and VDAC2, interacted with BVs, as determined with far-western blotting and reverse far-western blotting. We speculated that the proteins interacting with the virus could either enhance or inhibit the infection of the virus. The data provide comprehensive useful information for further research on the interaction of the host with BmNPV.
Insights
Researchers identified 24 proteins in silkworm midguts that bind to Bombyx mori nucleopolyhedrovirus (BmNPV). These proteins, involved in viral transport and metabolism, show varied responses to BmNPV infection, offering insights into silkworm resistance mechanisms.
Area of Science:
- * Insect Pathology
- * Molecular Biology
- * Proteomics
Background:
- * Bombyx mori nucleopolyhedrovirus (BmNPV) causes significant economic losses in sericulture.
- * The molecular mechanisms underlying silkworm resistance to BmNPV and host-virus interactions are poorly understood.
Purpose of the Study:
- * To identify silkworm midgut proteins that interact with BmNPV.
- * To investigate the role of these proteins in silkworm resistance to BmNPV infection.
Main Methods:
- * In vitro analysis of silkworm midgut subcellular fractions using 1D/2D electrophoresis and far-western blotting.
- * Mass spectrometry (MS) for protein identification.
- * Far-western blotting and reverse far-western blotting for validating protein-virus interactions.
Main Results:
- * 24 proteins specifically bound to BmNPV budded viruses (BVs) in mitochondrial and microsomal fractions.
- * Identified proteins are involved in viral transport, energy metabolism, apoptosis, and viral propagation.
- * Protein expression profiles varied across different resistant silkworm strains following BmNPV infection; most were downregulated in the A35 strain.
- * No BmNPV-binding proteins were found in the cytosolic fraction.
- * RACK1 and VDAC2 were confirmed as interacting proteins with BVs.
Conclusions:
- * The study identified key host proteins involved in BmNPV-silkworm interactions.
- * These interacting proteins may modulate BmNPV infection, either enhancing or inhibiting it.
- * The findings provide valuable data for further research into silkworm-BmNPV molecular interactions and resistance mechanisms.
