title: Subcellular localizations and time-course expression of dengue envelope and non-structural 1 proteins in human endothelial cells.
abstract: 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.
authors.affiliation: Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Jesdaporn
authors.initials: J
authors.lastname: Poungsawai
authors.forename: Rattiyaporn
authors.initials: R
authors.lastname: Kanlaya
authors.forename: Sa-nga
authors.initials: SN
authors.lastname: Pattanakitsakul
authors.forename: Visith
authors.initials: V
authors.lastname: Thongboonkerd
mesh_terms: D015153:Blotting, Western; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry; D002467:Cell Nucleus; D003599:Cytoskeleton / Q000737:chemistry; D003716:Dengue Virus / Q000472:pathogenicity*; D042783:Endothelial Cells / Q000821:virology*; D020869:Gene Expression Profiling; D006801:Humans; D007425:Intracellular Membranes / Q000737:chemistry; D015388:Organelles / Q000737:chemistry; D013997:Time Factors; D014759:Viral Envelope Proteins / Q000096:biosynthesis*; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis*
chemical_list: C051821:E-glycoprotein, Dengue virus type 2; C079340:NS1 protein, Dengue virus type 2; D014759:Viral Envelope Proteins; D017361:Viral Nonstructural Proteins