Surface proteins of C6/36 cells involved in dengue virus 4 binding and entry

Surface proteins of C6/36 cells involved in dengue virus 4 binding and entry Dengue virus (DENV) is the causative agent of the most important mosquito-borne viral disease, which is endemic to over 100 countries in tropical and subtropical areas of the world. It is transmitted to humans by Aedes mosquitoes. The first step in the viral infection of host cells is virion attachment to the plasma membrane, which is mediated by specific surface molecules. There are several molecules that participate in DENV infection of mosquitoes, but only a few have been identified. In this work, we co-purified 4 proteins from C6/36 cells using a recombinant DENV 4 E protein and identified them as 70 kDa Heat Shock and 70 kDa Heat Shock cognate proteins (HSP70/HSc70), Binding immunoglobulin protein (BiP), Thioredoxin/protein disulphide isomerase (PDI), and 44 kDa Endoplasmic reticulum resident protein (ERp44) via matrix-assisted laser desorption/ionisation time of flight (Maldi-ToF) analysis. Using immunofluorescence and flow cytometry assays, we observed re-localisation of HSP70/HSc70 and, to a lesser extent, BiP to the plasma membrane under stress conditions, such as during DENV infection. By performing binding and infection assays independently, we found that all 4 proteins participate in both processes, but to differing extents: HSP70/HSc70 is the most critical component, while ERp44 is less important. Viral infection was not inhibited when the cells were incubated with antibodies against all of the surface proteins after virus binding, which suggests that DENV entry to C6/36 cells is mediated by these proteins at the same step and not sequentially. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Surface proteins of C6/36 cells involved in dengue virus 4 binding and entry

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Publisher
Springer Vienna
Copyright
Copyright © 2013 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-012-1596-0
Publisher site
See Article on Publisher Site

Abstract

Dengue virus (DENV) is the causative agent of the most important mosquito-borne viral disease, which is endemic to over 100 countries in tropical and subtropical areas of the world. It is transmitted to humans by Aedes mosquitoes. The first step in the viral infection of host cells is virion attachment to the plasma membrane, which is mediated by specific surface molecules. There are several molecules that participate in DENV infection of mosquitoes, but only a few have been identified. In this work, we co-purified 4 proteins from C6/36 cells using a recombinant DENV 4 E protein and identified them as 70 kDa Heat Shock and 70 kDa Heat Shock cognate proteins (HSP70/HSc70), Binding immunoglobulin protein (BiP), Thioredoxin/protein disulphide isomerase (PDI), and 44 kDa Endoplasmic reticulum resident protein (ERp44) via matrix-assisted laser desorption/ionisation time of flight (Maldi-ToF) analysis. Using immunofluorescence and flow cytometry assays, we observed re-localisation of HSP70/HSc70 and, to a lesser extent, BiP to the plasma membrane under stress conditions, such as during DENV infection. By performing binding and infection assays independently, we found that all 4 proteins participate in both processes, but to differing extents: HSP70/HSc70 is the most critical component, while ERp44 is less important. Viral infection was not inhibited when the cells were incubated with antibodies against all of the surface proteins after virus binding, which suggests that DENV entry to C6/36 cells is mediated by these proteins at the same step and not sequentially.

Journal

Archives of VirologySpringer Journals

Published: Jun 1, 2013

References

  • Flavivirus genome organization, expression, and replication
    Chambers, TJ; Hahn, CS; Galler, R; Rice, CM
  • Dengue-2-virus-interacting polypeptides involved in mosquito cell infection
    Paingankar, MS; Gokhale, MD; Deobagkar, DN
  • Dengue viruses and mononuclear phagocytes. I. Infection enhancement by non-neutralizing antibody
    Halstead, SB; O’Rourke, EJ
  • Dendritic-cell-specific ICAM3-grabbing non-integrin is essential for the productive infection of human dendritic cells by mosquito-cell-derived dengue viruses
    Navarro-Sánchez, E; Almeyer, R; Schwartz, O; Fieschi, F; Virelizier, JL; Arenzana-Seisdedos, F; Desprès, P
  • Identification of GRP 78 (BiP) as a liver cell expressed receptor element for dengue virus serotype 2
    Jindadamrongwech, S; Thepparit, C; Smith, DR
  • Identification of dengue virus binding proteins using affinity chromatography
    Upanan, S; Kuadkikan, A; Smith, DR
  • Heat shock effect upon dengue virus replication into U937 cells
    Chavez-Salinas, S; Ceballos-Olvera, I; Reyes-del Valle, J; Medina, F; Angel, RM

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