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RAB4B and Japanese encephalitis virus E protein interaction is essential for viral entry in early endosomes

Chang Miao1, Qin Zhao2, Ya-Ting Zhang1

  • 1Research Center for Swine Diseases, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, China.

International Journal of Biological Macromolecules
|February 28, 2025
PubMed

Insights

Ras-Related GTP-Binding Protein 4b (RAB4B) is crucial for Japanese encephalitis virus (JEV) entry and replication. This study reveals RAB4B directly interacts with the JEV E protein, facilitating viral internalization and propagation.

Area of Science:

  • Virology
  • Cell Biology
  • Molecular Mechanisms

Background:

  • Ras-Related GTP-Binding Protein 4b (RAB4B) regulates intracellular trafficking.
  • Previous studies indicated RAB4B enhances Japanese encephalitis virus (JEV) replication.
  • The precise role of RAB4B in JEV internalization was not fully understood.

Purpose of the Study:

  • To elucidate the mechanisms by which RAB4B influences JEV replication.
  • To identify host factors mediating JEV entry.
  • To validate RAB4B as a key factor in JEV infection.

Main Methods:

  • Genome-wide CRISPR/Cas9 screening to identify host factors.
  • In vitro validation using RAB4B knockout and reintroduction in cell lines (U251, BV2).
  • Co-immunoprecipitation and binding assays to assess protein interactions.
  • In vivo studies using RAB4B knockout mice.

Main Results:

  • CRISPR/Cas9 screen identified RAB4B as essential for JEV replication.
  • RAB4B knockout cells exhibited significantly reduced JEV genome copies and viral titers.
  • RAB4B directly interacts with the JEV E protein.
  • RAB4B is necessary for JEV internalization into early endosomes.
  • RAB4B knockout mice showed reduced viral brain load and increased survival.

Conclusions:

  • RAB4B is indispensable for JEV entry into host cells.
  • The interaction between RAB4B and JEV E protein is critical for viral replication.
  • RAB4B represents a potential therapeutic target for JEV infection.
Keywords:
CRISPR/Cas9 screenJapanese encephalitis virus E proteinsRas-related GTP-binding protein 4bViral entry

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