title: Identification and characterization of a 43 kDa actin protein involved in the DENV-2 binding and infection of ECV304 cells.
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.
authors: Department of Microbiology, College of Basic Medical Sciences, Third Military Medical University, Key Laboratory of Microbial Engineering under the Educational Committee in Chongqing, Chongqing 400038, PR China.
authors: Jie
authors: J
authors: Yang
authors: Lingyun
authors: L
authors: Zou
authors: Zhen
authors: Z
authors: Hu
authors: Wei
authors: W
authors: Chen
authors: Junlei
authors: J
authors: Zhang
authors: Junmin
authors: J
authors: Zhu
authors: Xin
authors: X
authors: Fang
authors: Wenchang
authors: W
authors: Yuan
authors: Xiaomei
authors: X
authors: Hu
authors: Fuquan
authors: F
authors: Hu
authors: Xiancai
authors: X
authors: Rao
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D002460:Cell Line; D003716:Dengue Virus / Q000502:physiology*; D004847:Epithelial Cells / Q000821:virology*; D054884:Host-Pathogen Interactions; D006801:Humans; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D011485:Protein Binding; D025941:Protein Interaction Mapping; D014759:Viral Envelope Proteins / Q000378:metabolism*; D053586:Virus Internalization
chemical_list: D000199:Actins; C051821:E-glycoprotein, Dengue virus type 2; D014759:Viral Envelope Proteins