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A hypothetical model of crossing Bombyx mori nucleopolyhedrovirus through its host midgut physical barrier.

Cheng Y, Wang XY, Hu H, Killiny N, Xu JP - PLoS ONE (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Anhui Agricultural University, Hefei, People's Republic of China.

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.

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Related in: MedlinePlus

Virus binding experiment on total proteins of B. mori midgut resolved by 2-DE.A) Virus overlay binding experiment. B) Separation of B. mori midgut total proteins by 2-DE. Molecular mass was indicated on the left and isoelectric point (PI) range on the top. Arrows named A to H in A) and B) referred to the detected spots on PVDF membrane and the corresponding proteins in gel, respectively.
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pone-0115032-g003: Virus binding experiment on total proteins of B. mori midgut resolved by 2-DE.A) Virus overlay binding experiment. B) Separation of B. mori midgut total proteins by 2-DE. Molecular mass was indicated on the left and isoelectric point (PI) range on the top. Arrows named A to H in A) and B) referred to the detected spots on PVDF membrane and the corresponding proteins in gel, respectively.

Mentions: Total proteins were also characterized by 2-DE followed by far-Western blot experiment, and eight spots were visualized clearly and indicated by arrows named A to H respectively (Fig. 3A). The eight corresponding spots in gel were also indicated by arrows in Fig. 3B and cut out for MALDI-TOF/TOF MS analysis.


A hypothetical model of crossing Bombyx mori nucleopolyhedrovirus through its host midgut physical barrier.

Cheng Y, Wang XY, Hu H, Killiny N, Xu JP - PLoS ONE (2014)

Virus binding experiment on total proteins of B. mori midgut resolved by 2-DE.A) Virus overlay binding experiment. B) Separation of B. mori midgut total proteins by 2-DE. Molecular mass was indicated on the left and isoelectric point (PI) range on the top. Arrows named A to H in A) and B) referred to the detected spots on PVDF membrane and the corresponding proteins in gel, respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4264868&req=5

pone-0115032-g003: Virus binding experiment on total proteins of B. mori midgut resolved by 2-DE.A) Virus overlay binding experiment. B) Separation of B. mori midgut total proteins by 2-DE. Molecular mass was indicated on the left and isoelectric point (PI) range on the top. Arrows named A to H in A) and B) referred to the detected spots on PVDF membrane and the corresponding proteins in gel, respectively.
Mentions: Total proteins were also characterized by 2-DE followed by far-Western blot experiment, and eight spots were visualized clearly and indicated by arrows named A to H respectively (Fig. 3A). The eight corresponding spots in gel were also indicated by arrows in Fig. 3B and cut out for MALDI-TOF/TOF MS analysis.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Anhui Agricultural University, Hefei, People's Republic of China.

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.

Show MeSH
Related in: MedlinePlus