The Ubiquitin-Proteasome Pathway Is Important for Dengue Virus Infection in Primary Human Endothelial Cells
Resource
JOURNAL OF PROTEOME RESEARCH, 9(10), 4960-4971
Journal
JOURNAL OF PROTEOME RESEARCH
Journal Volume
9
Journal Issue
10
Pages
4960-4971
Date Issued
2010
Date
2010
Author(s)
Kanlaya, Rattiyaporn
Pattanakitsakul, Sa-nga
Sinchaikul, Supachok
Chen, Shui-Tein
Thongboonkerd, Visith
Abstract
Dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are the most severe forms of dengue virus infection with hemorrhage and plasma leakage. However, pathogenic mechanisms of DHF and DSS remain poorly understood. We therefore investigated host responses as determined by changes in the cellular proteome of primary human endothelial cells upon infection with dengue virus serotype 2 (DEN-2) at a multiplicity of infection (MOI) of 10 for 24 h. Two-dimensional PAGE and quantitative intensity analysis revealed 38 significantly altered protein spots (16 upregulated and 22 downregulated) in DEN-2-infected cells compared to mock controls. These altered proteins were successfully identified by mass spectrometry, including those involved in oxidative stress response, transcription and translation, cytoskeleton assembly, protein degradation, cell growth regulation, apoptosis, cellular metabolism, and antiviral response. The proteomic data were validated by Western blot analyses [upregulated ubiquitin-activating enzyme E1 (UBE1) and downregulated annexin A2] and an immunofluorescence study (upregulated MxA). Interestingly, we found that MxA was colocalized with DEN-2 viral capsid protein, strengthening its role as an antiviral protein. Moreover, we also identified upregulation of a proteasome subunit. Our functional study revealed the significant role of ubiquitination in dengue infection and UBE1 inhibition by its specific inhibitor (UBEI-41) caused a significant reduction in the level of viral protein synthesis and its infectivity. Our findings suggest that various biological processes were triggered in response to dengue infection, particularly antiviral IFN and ubiquitin-proteasome pathways. ? 2010 American Chemical Society.
Subjects
Dengue; proteasome; proteomics; therapeutic targets; ubiquitin
SDGs
Other Subjects
capsid protein; lipocortin 2; proteome; ubiquitin protein ligase E3; virus protein; apoptosis; article; cell growth; cell metabolism; cell viability; controlled study; cytoskeleton; dengue; Dengue virus 2; down regulation; endothelium cell; enzyme inhibition; genetic transcription; human; human cell; immunofluorescence; mass spectrometry; oxidative stress; polyacrylamide gel electrophoresis; priority journal; protein degradation; protein expression; protein localization; protein synthesis; proteomics; quantitative analysis; RNA translation; ubiquitination; upregulation; Western blotting; Annexin A2; Blotting, Western; Capsid Proteins; Cells, Cultured; Dengue Virus; Electrophoresis, Gel, Two-Dimensional; Endothelial Cells; GTP-Binding Proteins; Host-Pathogen Interactions; Humans; Proteasome Endopeptidase Complex; Proteomics; Signal Transduction; Ubiquitin; Ubiquitin-Activating Enzymes; Dengue virus; Miridae
Type
journal article
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