Inhibition of orf virus replication in goat skin fibroblast cells by the HSPA1B protein, as demonstrated by iTRAQ-based
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ORIGINAL ARTICLE
Inhibition of orf virus replication in goat skin fibroblast cells by the HSPA1B protein, as demonstrated by iTRAQ‑based quantitative proteome analysis Jun‑hong Hao1 · Han‑jin Kong1 · Ming‑hao Yan1 · Chao‑chao Shen1 · Guo‑wei Xu1 · Da‑jun Zhang1 · Ke‑shan Zhang1 · Hai‑xue Zheng1 · Xiang‑tao Liu1 Received: 8 March 2020 / Accepted: 26 July 2020 © Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract Orf virus (ORFV) infects sheep and goat tissues, resulting in severe proliferative lesions. To analyze cellular protein expression in ORFV-infected goat skin fibroblast (GSF) cells, we used two-dimensional liquid chromatography-tandem mass spectrometry coupled with isobaric tags for relative and absolute quantification (iTRAQ). The proteomics approach was used along with quantitative reverse transcription polymerase chain reaction (RT-qPCR) to detect differentially expressed proteins in ORFV-infected GSF cells and mock-infected GSF cells. A total of 282 differentially expressed proteins were identified. It was found that 222 host proteins were upregulated and 60 were downregulated following viral infection. We confirmed that these proteins were differentially expressed and found that heat shock 70-kDa protein 1B (HSPA1B) was differentially expressed and localized in the cytoplasm. It was also noted that HSPA1B caused inhibition of viral proliferation, in the middle and late stages of viral infection. The differentially expressed proteins were associated with the biological processes of viral binding, cell structure, signal transduction, cell adhesion, and cell proliferation.
Introduction Orf, which is caused by orf virus (ORFV), is one of the most widespread viral diseases worldwide. Although ORFV mainly infects sheep and goats, it can also infect other ruminants and other mammals [18]. Genes involved in virulence and pathogenesis are distributed in the ITR regions of the ORFV genome. The virus encodes involved in immunomodulation, including viral-interleukin 10 (vIL‐10, ORFV127), vascular endothelial growth factor (VEGF, ORFV132), orf virus interferon resistance protein (OVIFNR, ORFV020), chemokine binding protein (CBP, ORFV112), granulocyte–macrophage colony-stimulating factor/interleukin-2 (GM-CSF/IL-2) inhibitory protein (GIF, ORFV117), nuclear Handling Editor: William G Dundon. * Ke‑shan Zhang [email protected] 1
State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute of Chinese Academy of Agriculture Science, No. 1, Xujiaping, Lanzhou 730046, Gansu, People’s Republic of China
factor kappa B (NF‐κB) inhibitory protein (ORFV125), and deoxyuridine 5′-triphosphate pyrophosphatase (dUTPase, ORFV007) [38]. Like other poxviruses, ORFV encodes a range of molecules that play vital roles in immune evasion by the production of anti-inflammatory proteins [13]. These proteins, which are mainly involved in the interaction with host defense mechanisms, include OVIFNR, GIF, vIL-10, and VEGF. A number o
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