Subcellular localizations and time-course expression of dengue envelope and non-structural 1 proteins in human endothelial cells

• Published in: Microbial pathogenesis Vol. 51; no. 3; pp. 225 - 229
• Main Authors: Poungsawai, Jesdaporn, Kanlaya, Rattiyaporn, Pattanakitsakul, Sa-nga, Thongboonkerd, Visith
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier India Pvt Ltd 01.09.2011; Elsevier
• Subjects: Biological and medical sciences; Blotting, Western; Cell Fractionation; Cell Membrane - chemistry; Cell Nucleus; cytoskeleton; Cytoskeleton - chemistry; cytosol; Dengue; Dengue virus; Dengue Virus - pathogenicity; Endothelial cells; Endothelial Cells - virology; Envelope; fluorescent antibody technique; fractionation; Fundamental and applied biological sciences. Psychology; Gene Expression Profiling; Humans; Intracellular Membranes - chemistry; Microbiology; NS1; Organelles - chemistry; proteins; Subcellular localization; Time Factors; Viral Envelope Proteins - biosynthesis; Viral Nonstructural Proteins - biosynthesis; Viral proteins; Western blotting; RT; E; Dengue; MOI; Envelope; DEN-2; NS1; ER; hnRNP C1/C2; Endothelial cells; BSA; Viral proteins; FBS; Subcellular localization; Human; Endothelial cell; Dengue 1; Non structural protein; Infection; Viral disease; Arbovirus disease; Localization
• ISSN: 0882-4010; 1096-1208
• DOI: 10.1016/j.micpath.2011.04.011

We performed subcellular fractionation of human endothelial cells (EA.hy926) after a challenge with dengue virus serotype-2 (DEN-2) at multiplicity of infection (MOI) of 10 for 2 h. The cells were fractionated into four fractions including cytosol, membrane/organelle, nucleus and cytoskeleton, and their purity was nicely confirmed by immunofluorescence staining of markers for individual subcellular compartments. Western blot analysis of dengue envelope (E) and non-structural 1 (NS1) proteins showed that E was present only in membrane/organelle fraction, whereas NS1 was present in membrane/organelle, nucleus, and cytoskeleton fractions. Time-course study showed that E was present immediately after the challenge, whereas NS1 was present after 12 h of the challenge. Our data may provide a new roadmap for studying mechanisms of replication of dengue virus and its interactions with host cells.