Endocytic pathway followed by dengue virus to infect the mosquito cell line C6/36 HT

Virology. 2008 Aug 15;378(1):193-9. doi: 10.1016/j.virol.2008.05.012. Epub 2008 Jun 20.

Abstract

The endocytic pathway followed by dengue virus to infect the mosquito cells C6/36 HT was analyzed. Using DIL-labeled virions and real-time imaging it was determined that viral entry into C6/36 HT takes approximately 5 to 7 min. Pretreatment of C6/36 HT cells with sucrose and bafilomycin A, but not filipin, inhibited dengue virus infection up to 80%. Furthermore, the overexpression of dominant-negative mutants of Eps15, a molecule required for the formation of clathrin-coated vesicles, reduced dengue infection up to 50%, indicating that dengue virus entry is through clathrin-mediated endocytosis and is pH-dependent. By double-immunofluorescence assays, DIL-labeled particles were colocalized with early endosomes at 5 min and with lysosomes mainly at 30 min post-infection. Finally, disruption of the microtubule and microfilaments by nocodazole and by cytochalasin D reduced viral infection by more than 80%. Taken together these results indicate that dengue virions enter into C6/36 HT cells by clathrin-mediated endocytosis, using the endosomal pathway from early endosomes to acidic lysosomes before viral RNA is released into the cytoplasm.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aedes / classification
  • Aedes / cytology*
  • Aedes / virology*
  • Animals
  • Cell Line
  • Cells, Cultured
  • Clathrin / metabolism*
  • Cricetinae
  • Dengue Virus / genetics
  • Dengue Virus / metabolism
  • Dengue Virus / pathogenicity*
  • Endocytosis / physiology*
  • Endosomes / virology
  • Fluorescent Antibody Technique
  • Lysosomes / virology
  • Microscopy, Confocal
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism
  • Virion / pathogenicity
  • Virus Internalization

Substances

  • Clathrin
  • Viral Nonstructural Proteins