Identification of regulators of the early stage of XXXX virus infection during curcumin treatment.

@entity14595 @entity4404 @entity1457 @entity9487 @entity1093 @entity19330 @entity10282 @entity14875 @entity8946 @entity42696
The effect of curcumin pretreatment (15-240   M) in fathead minnow cells infected with @entity14595 virus (VHSV) was evaluated. Cell viability, apoptosis and viral copy number were analyzed using Cell Counting Kit-8 assay, @entity4404 staining, and reverse transcription-PCR, respectively. Pretreatment with 120   M curcumin showed an increase in viability (>90% of mock) of VHSV-infected cells and reduction in the copy number (0.2-log reduction in VHSV N gene expression), reactive oxygen species and apoptosis in the cells without cytotoxic effects. To understand the mechanisms underlaying the antiviral effects of curcumin pretreatment, a comparative proteomic analysis was performed in four samples (M, mock; C, curcumin-treated; V, VHSV-infected; and CV, curcumin-treated VHSV-infected) in triplicate. In total, 185 proteins were detected. The analysis showed that three proteins, including @entity1457 cognate 71 ( @entity9487 ), actin, alpha cardiac muscle ( @entity42696 ) and elongation factor 1 ( @entity1093 ) were differentially expressed between V and CV samples. Network analysis performed by Ingenuity Pathways Analysis (IPA) showed that @entity9487 was the primary protein interacting with fibronectin (FN) 1, actins ( @entity14875 , @entity10282 , F-actin) and @entity19330 (GSN) in both V and CV samples and thus is a strong target candidate for the protection from @entity14595 at the viral entry stage. Our proteomics data suggest that curcumin pretreatment inhibits entry of VHSV in cells by downregulating @entity8946 or upregulating F-actin. For both proteins, @entity9487 acts as a binding protein that modulates their functions. Furthermore, consistent with the effect of a @entity1457 protein inhibitor (KNK437), curcumin downregulated @entity9487 expression with increasing viability of VHSV-infected cells and inhibited VHSV replication, suggesting that the downregulation of @entity9487 could be responsible for the antiviral activity of curcumin. In conclusion, this study indicates that the suppression of viral entry by rearrangement of the F-actin/G-actin ratio via downregulating @entity9487 is a plausible mechanism by which curcumin pretreatment controls the early stages of @entity14595 .
