Identification and characterization of a 43 kDa actin protein involved in the DENV-2 binding and infection of ECV304 cells |
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| Institution: | 1. Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, Av. Universidad 3000, Ciudad Universitaria, Coyoacán, 04510, Ciudad de México, Mexico;2. Departamento de Biomedicina Molecular, Centro de Investigación y de Estudios Avanzados del IPN, Av. Instituto Politécnico Nacional 2508, Gustavo A. Madero, San Pedro Zacatenco, 07360, Ciudad de México, Mexico;1. Center for Health Disparities and Molecular Medicine, Department of Basic Sciences, Loma Linda, University School of Medicine, Loma Linda, CA, 92354, USA;2. Division of Biochemistry, Department of Basic Sciences, Loma Linda, University School of Medicine, Loma Linda, CA, 92354, USA;3. University of Texas at San Antonio, San Antonio, TX, USA;4. Section of Endocrinology, Riverside University Health System Medical Center, Moreno Valley, CA, USA;1. Department of Aquaculture, Faculty of Natural Resources and Marine Sciences, Tarbiat Modares University, 64414-356, Noor, Iran;2. Department of Nanobiotechnology, Faculty of Biological Science, Tarbiat Modares University, Tehran, Iran;1. Department of Cellular Neurobiology, Graduate School of Medicine, The University of Tokyo, Tokyo 113-0033, Japan;2. Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto 606-8507, Japan |
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| Abstract: | Characterization of the primary host factors associated with host–virus interaction is critical for understanding how a virus infects its host cell. In this study, a modified virus overlay protein binding assay was developed. Host factors with 34, 43, and 55 kDa proteins, which could interact with EDIII, a cell receptor-binding domain of Dengue virus (DENV)-enveloped E protein, were isolated from ECV304 cells. Mass spectrometry identified peptide masses of 43 kDa protein matched to actin, a cytoskeleton protein in eukaryotic cells. The interaction between 43 kDa actin and DENV-2 EDIII was further confirmed by competitive blocking and co-immunoprecipitation assays. Actin cytoskeleton rearrangement was observed within 1 h p.i. of DENV-2-infected ECV304 cells in the confocal immunofluorescent assay. The co-localization of DENV-2 E protein with the actin filaments occurred in the late stage of the DENV replication cycle. Finally, a docking complex was constructed, and the functional residues involved in the interaction of actin and DENV-2 EDIII protein were predicted. Our findings suggest that the direct contact of DENV E protein with 43 kDa actin protein may have a crucial function in DENV infection of ECV304 cells. |
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