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Bovine ephemeral fever virus uses a clathrin-mediated and dynamin 2-dependent endocytosis pathway that requires Rab5

Ching Y Cheng1, Wing L Shih, Wei R Huang

  • 1Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan.

Journal of Virology
|October 12, 2012
PubMed

Insights

Bovine ephemeral fever virus (BEFV) enters cells via clathrin-mediated endocytosis, a process dependent on dynamin 2, early/late endosomes, and microtubules. This pH-dependent pathway is crucial for viral infection and replication.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Biology

Background:

  • The cellular entry mechanisms of bovine ephemeral fever virus (BEFV) remain largely uncharacterized.
  • Understanding viral entry pathways is critical for developing antiviral strategies.

Purpose of the Study:

  • To elucidate the specific cell pathways utilized by BEFV during host cell entry.
  • To investigate the roles of clathrin, dynamin 2, endosomes, and microtubules in BEFV infection.

Main Methods:

  • Fluorescence microscopy to observe M protein colocalization with clathrin and dynamin 2.
  • Internalization studies using fluorescently labeled BEFV (DiO).
  • Genetic and pharmacological inhibition of endocytosis-related proteins (Rab5, Rab7a, microtubules) and endosomal acidification.

Main Results:

  • BEFV M protein colocalized with clathrin and dynamin 2, indicating involvement in endocytosis.
  • BEFV internalization was confirmed to be clathrin-mediated and dynamin 2-dependent.
  • BEFV infection required both early (Rab5) and late (Rab7a) endosomes.
  • Microtubule disruption (nocodazole) reduced viral M protein synthesis and yield.
  • Endosomal acidification inhibitors blocked BEFV infection, confirming a pH-dependent entry process.

Conclusions:

  • BEFV primarily enters host cells through clathrin-mediated endocytosis, a pathway reliant on dynamin 2.
  • The virus utilizes both early and late endosomes, with microtubules facilitating intracellular trafficking.
  • Endosomal acidification is essential for successful BEFV entry and subsequent infection.

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