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

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 t...

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Published inMicrobial pathogenesis Vol. 51; no. 3; pp. 225 - 229
Main Authors Poungsawai, Jesdaporn, Kanlaya, Rattiyaporn, Pattanakitsakul, Sa-nga, Thongboonkerd, Visith
Format Journal Article
LanguageEnglish
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
Online AccessGet full text

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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.
AbstractList 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.
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.
Author Thongboonkerd, Visith
Pattanakitsakul, Sa-nga
Poungsawai, Jesdaporn
Kanlaya, Rattiyaporn
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Issue 3
Keywords 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
Language English
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Snippet We performed subcellular fractionation of human endothelial cells (EA.hy926) after a challenge with dengue virus serotype-2 (DEN-2) at multiplicity of...
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SubjectTerms 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
Title Subcellular localizations and time-course expression of dengue envelope and non-structural 1 proteins in human endothelial cells
URI https://dx.doi.org/10.1016/j.micpath.2011.04.011
https://www.ncbi.nlm.nih.gov/pubmed/21575706
https://search.proquest.com/docview/873494901
https://search.proquest.com/docview/904480250
Volume 51
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