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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: 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.

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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Real-time PCR analysis of expression profiles of BmNPV binding proteins in B. mori midguts.Columns with different colors indicated the experimental treats to the larvae. A to L refer to relative expression level of ATP-A, ATP-B, AK, RP0, Actin, En, PGK, GDAP, PHB2, HSP60, Crt, and RCX1 respectively. Data were normalized usingBmrps3and represented as means±standard errors of the means from three independent experiments. ** Indicates statistical significance<0.01(ANOVA and LSD aposteriori test).
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pone-0115032-g004: Real-time PCR analysis of expression profiles of BmNPV binding proteins in B. mori midguts.Columns with different colors indicated the experimental treats to the larvae. A to L refer to relative expression level of ATP-A, ATP-B, AK, RP0, Actin, En, PGK, GDAP, PHB2, HSP60, Crt, and RCX1 respectively. Data were normalized usingBmrps3and represented as means±standard errors of the means from three independent experiments. ** Indicates statistical significance<0.01(ANOVA and LSD aposteriori test).

Mentions: Real-time qPCR analysis was performed between BmNPV infected P50 larvae and the control group treated with ddH2O. Fifth instar molt larvae of P50 were starved overnight and fed with 500 OB of BmNPV T3 strain or ddH2O per larva orally. The relative expression levels of PGK (Fig. 4G) and GDAP (Fig. 4H) in P50 midguts were up-regulated significantly (P<0.01) at 48hpi, while the levels of eight genes, ATP-A (Fig. 4A), AK (Fig. 4C), RP0 (Fig. 4D), actin (Fig. 4E), En (Fig. 4F), PHB2 (Fig. 4I), HSP (Fig. 4J), Crt (Fig. 4K), respectively, were down-regulated significantly (P<0.01) at 48hpi. The relative expression levels of ATP-B (Fig. 4B) and RCX1 (Fig. 4L) were not significantly different between the infected and the control.


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)

Real-time PCR analysis of expression profiles of BmNPV binding proteins in B. mori midguts.Columns with different colors indicated the experimental treats to the larvae. A to L refer to relative expression level of ATP-A, ATP-B, AK, RP0, Actin, En, PGK, GDAP, PHB2, HSP60, Crt, and RCX1 respectively. Data were normalized usingBmrps3and represented as means±standard errors of the means from three independent experiments. ** Indicates statistical significance<0.01(ANOVA and LSD aposteriori test).
© Copyright Policy
Related In: Results  -  Collection

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

pone-0115032-g004: Real-time PCR analysis of expression profiles of BmNPV binding proteins in B. mori midguts.Columns with different colors indicated the experimental treats to the larvae. A to L refer to relative expression level of ATP-A, ATP-B, AK, RP0, Actin, En, PGK, GDAP, PHB2, HSP60, Crt, and RCX1 respectively. Data were normalized usingBmrps3and represented as means±standard errors of the means from three independent experiments. ** Indicates statistical significance<0.01(ANOVA and LSD aposteriori test).
Mentions: Real-time qPCR analysis was performed between BmNPV infected P50 larvae and the control group treated with ddH2O. Fifth instar molt larvae of P50 were starved overnight and fed with 500 OB of BmNPV T3 strain or ddH2O per larva orally. The relative expression levels of PGK (Fig. 4G) and GDAP (Fig. 4H) in P50 midguts were up-regulated significantly (P<0.01) at 48hpi, while the levels of eight genes, ATP-A (Fig. 4A), AK (Fig. 4C), RP0 (Fig. 4D), actin (Fig. 4E), En (Fig. 4F), PHB2 (Fig. 4I), HSP (Fig. 4J), Crt (Fig. 4K), respectively, were down-regulated significantly (P<0.01) at 48hpi. The relative expression levels of ATP-B (Fig. 4B) and RCX1 (Fig. 4L) were not significantly different between the infected and the control.

Bottom Line: 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.

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