Identification and characterization of a 43 kDa actin protein involved in the DENV-2 binding and infection of ECV304 cells

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 E...

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Bibliographic Details
Published in:Microbes and infection 2013-04, Vol.15 (4), p.310-318
Main Authors: Yang, Jie, Zou, Lingyun, Hu, Zhen, Chen, Wei, Zhang, Junlei, Zhu, Junmin, Fang, Xin, Yuan, Wenchang, Hu, Xiaomei, Hu, Fuquan, Rao, Xiancai
Format: Article
Language:English
Subjects:
Actin
Actins - chemistry
Actins - metabolism
Cell Line
Dengue virus
Dengue Virus - physiology
ECV304 cells
EDIII protein
Epithelial Cells - virology
Host-Pathogen Interactions
Humans
Mass Spectrometry
Microscopy, Confocal
Microscopy, Fluorescence
Molecular Weight
Protein Binding
Protein Interaction Mapping
Viral Envelope Proteins - metabolism
Virus Internalization
VOPBA
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Summary: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.
ISSN:1286-4579
1769-714X
DOI:10.1016/j.micinf.2013.01.004