Project description:Ubiquitin-fold modifier 1 (UFM1) is attached to protein substrates through the sequential activity of an E1 (UBA5)-E2 (UBC1)-E3 (UBL1) cascade. UFBP1 is a conserved UFL1-interacting protein in mammals. However, there is no study on UFBP1 in silkworm yet. In this study, we identified a UFBP1 ortholog in B. mori genome. Spatio-Temporal expression profiles showed that BmUFBP1 expression was highly in midgut, fatbody, and at the moth stage. BmUFBP1 knockdown inhibited ER-stress induced ER chaperone expression and BmNPV proliferation, while BmUFBP1 overexpression increased BmNPV proliferation. Furthermore, RNA sequencing and experimental verification identified the HSP70 family member BmBIP as the BmUFBP1 downstream target in regulating BmNPV proliferation. Overall, these results suggest that BmUFBP1 facilitates BmNPV proliferation via ATF6-BIP signaling.
Project description:Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the most acute infectious diseases in silkworm, which has caused great economic loss in sericulture. Previous study showed that the content of components in mulberry leaves, particularly for moracin N, was increased after UV-B irradiation. In this study, the BmNPV resistance of silkworms reared on UV-B treated and moracin N spreaded mulberry leaves was improved. To uncover the mechanism of enhanced BmNPV resistance, silkworm midguts from UV-B treated mulberry leaves (BUM) and moracin N (BNM) groups were analyzed by SWATH-based proteomic technique. Of note, the abundance of ribosomal proteins in BUM and BNM groups was significantly changed to maintain the synthesis of total protein levels and cell survival. While, cytochrome c oxidase subunit II, calcium ATPase and programmed cell death 4 involved in apoptotic process were up-regulated in BNM group. Expressions of lipase-1, serine protease precursor, Rab1 protein, and histone genes were increased significantly in BNM group. These results suggest that moracin N might be the main active components in UV-B treated mulberry leaves to affect the BmNPV-resistance of silkworm, which could promote apoptotic cell death, enhance the organism immunity, and regulate the intercellular environment of cells in silkworm. It also presents an innovative process to reduce the mortality rate of silkworm infected with BmNPV.
Project description:The molecular mechanism involved in BmNPV resistance was investigated using a genome wide microarray in the midgut tissues of BmNPV infected resistant (Sarupat) and susceptible (CSR-2) Indian silkworm races. In the resistant race, 735 genes were upregulated and 589 genes were down regulated at 12 hours of BmNPV post infection. Similarly in case of susceptible race, 2183 genes were up regulated and 2115 genes were down regulated. Among the differentially expressed genes, nine upregulated and eight down regulated genes were validated using real time qPCR analysis. In Sarupat, the significantly upregulated genes are vacuolar protein sorting associated gene, X fin like protein and Carboxy peptidase E like protein in BmNPV infected midguts, whereas the prominent down regulated genes are glutamate receptor ionotropic kainite 2-like, BTB/POZ domain and transferrin. In the case of CSR-2, the considerably upregulated genes are Peptidoglycan recognition protein S6 precursor and rapamycin while the conspicuous down regulated genes are facilitated trehalose transporter and zinc transporter ZIP1-like gene. Our results provided a vital insights of Bombyx mori in reference with its molecular mechanism in immune response against the BmNPV invasion.
Project description:The molecular mechanism involved in BmNPV resistance was investigated using a genome wide microarray in the midgut tissues of BmNPV infected resistant (Sarupat) and susceptible (CSR-2) Indian silkworm races. In the resistant race, 735 genes were upregulated and 589 genes were down regulated at 12 hours of BmNPV post infection. Similarly in case of susceptible race, 2183 genes were up regulated and 2115 genes were down regulated. Among the differentially expressed genes, nine upregulated and eight down regulated genes were validated using real time qPCR analysis. In Sarupat, the significantly upregulated genes are vacuolar protein sorting associated gene, X fin like protein and Carboxy peptidase E like protein in BmNPV infected midguts, whereas the prominent down regulated genes are glutamate receptor ionotropic kainite 2-like, BTB/POZ domain and transferrin. In the case of CSR-2, the considerably upregulated genes are Peptidoglycan recognition protein S6 precursor and rapamycin while the conspicuous down regulated genes are facilitated trehalose transporter and zinc transporter ZIP1-like gene. Our results provided a vital insights of Bombyx mori in reference with its molecular mechanism in immune response against the BmNPV invasion. Organism : Bombyx mori , Agilent Custom Silkworm Gene Expression 4x44k Array (AMADID: 066047) designed by Genotypic Technology Private Limited.
Project description:Bombyx mori nucleopolyhedrovirus (BmNPV) relies on the envelope glycoprotein GP64 to mediate host cell entry. Intriguingly, both the wild-type GP64 retaining its signal peptide (SP) and the SP-cleaved variant (SP∆nGP64) can support viral invasion, yet their mechanisms may differ. To identify potential GP64 receptors and dissect host interaction profiles, we performed comparative co-immunoprecipitation using purified GP64 and SP∆nGP64, followed by data-independent acquisition (DIA)-based proteomic analysis. A total of 6,064 and 5,247 host proteins were identified, respectively. Enrichment analyses revealed shared involvement in membrane trafficking, lipid metabolism, and endocytosis—pathways essential for viral entry.
Project description:Employed BmN cell after 10 MOI (multiplicity of infection) BmNPV 36 hpi (hour post-infection) to compare the proteomics by LC-MS/MS