title: Identification of regulators of the early stage of viral hemorrhagic septicemia virus infection during curcumin treatment.
abstract: The effect of curcumin pretreatment (15-240 μM) in fathead minnow cells infected with viral hemorrhagic septicemia virus (VHSV) was evaluated. Cell viability, apoptosis and viral copy number were analyzed using Cell Counting Kit-8 assay, Annexin V staining, and reverse transcription-PCR, respectively. Pretreatment with 120 μM 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 heat shock cognate 71 (HSC71), actin, alpha cardiac muscle (ACTC1) and elongation factor 1 (EEF1) were differentially expressed between V and CV samples. Network analysis performed by Ingenuity Pathways Analysis (IPA) showed that HSC71 was the primary protein interacting with fibronectin (FN) 1, actins (ACTB, ACTG, F-actin) and gelsolin (GSN) in both V and CV samples and thus is a strong target candidate for the protection from VHSV infection at the viral entry stage. Our proteomics data suggest that curcumin pretreatment inhibits entry of VHSV in cells by downregulating FN1 or upregulating F-actin. For both proteins, HSC71 acts as a binding protein that modulates their functions. Furthermore, consistent with the effect of a heat shock protein inhibitor (KNK437), curcumin downregulated HSC71 expression with increasing viability of VHSV-infected cells and inhibited VHSV replication, suggesting that the downregulation of HSC71 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 HSC71 is a plausible mechanism by which curcumin pretreatment controls the early stages of VHSV infection.
authors: Department of Food Science and Technology, Chonnam National University, Gwangju 500-757, South Korea.
authors: Eun-Hye
authors: EH
authors: Jeong
authors: Department of Food Science and Technology, Chonnam National University, Gwangju 500-757, South Korea; Bioenergy Research Center, Chonnam National University, Gwangju 500-757, South Korea.
authors: Bipin
authors: B
authors: Vaidya
authors: Department of Food Science and Technology, Chonnam National University, Gwangju 500-757, South Korea.
authors: Se-Young
authors: SY
authors: Cho
authors: Aquatic Life Disease Control Division, National Fisheries Research and Development Institute, Busan 619-705, South Korea.
authors: Myoung-Ae
authors: MA
authors: Park
authors: Department of Food Science and Technology, Chonnam National University, Gwangju 500-757, South Korea.
authors: Kusuma
authors: K
authors: Kaewintajuk
authors: West Sea Fisheries Research Institute, National Fisheries Research and Development Institute, Incheon 400-420, South Korea.
authors: Seok Ryel
authors: SR
authors: Kim
authors: Department of Aqualife Medicine, Chonnam National University, Yeosu 550-749, South Korea.
authors: Myung-Joo
authors: MJ
authors: Oh
authors: Biological Disaster Analysis Group, Korea Basic Science Institute, Daejeon 305-806, South Korea.
authors: Jong-Soon
authors: JS
authors: Choi
authors: Biological Disaster Analysis Group, Korea Basic Science Institute, Daejeon 305-806, South Korea. Electronic address: joseph@kbsi.re.kr.
authors: Joseph
authors: J
authors: Kwon
authors: Department of Food Science and Technology, Chonnam National University, Gwangju 500-757, South Korea; Bioenergy Research Center, Chonnam National University, Gwangju 500-757, South Korea; Agribio Disaster Research Center, Institute of Environmentally-Friendly Agriculture, Chonnam National University, Gwangju 500-757, South Korea. Electronic address: dwkim@jnu.ac.kr.
authors: Duwoon
authors: D
authors: Kim
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000008:administration & dosage / Q000494:pharmacology*; D003474:Curcumin / Q000008:administration & dosage / Q000494:pharmacology*; D003530:Cyprinidae; D005393:Fish Diseases / Q000821:virology*; D015870:Gene Expression / Q000187:drug effects; D031941:Hemorrhagic Septicemia, Viral / Q000821:virology*; D029141:Novirhabdovirus / Q000187:drug effects* / Q000502:physiology; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D014764:Viral Proteins; D003474:Curcumin
keywords: Curcumin; Heat shock cognate 71; Viral hemorrhagic septicemia virus