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The C-terminal helical domain of dengue virus precursor membrane protein is involved in virus assembly and entry

Szu-Chia Hsieh1, Gang Zou, Wen-Yang Tsai

  • 1Institute of Microbiology, College of Medicine, National Taiwan University, Taipei, Taiwan.

Virology
|December 7, 2010
PubMed

Insights

Site-directed mutagenesis revealed that specific proline substitutions in the dengue virus (DENV) precursor membrane protein's α-helical domain are crucial for virus replication, affecting both particle assembly and viral entry.

Area of Science:

  • Virology
  • Molecular Biology
  • Structural Biology

Background:

  • The dengue virus (DENV) precursor membrane (prM) protein contains an α-helical domain (MH) essential for viral replication.
  • Understanding the specific roles of different regions within the MH domain is critical for developing antiviral strategies.

Purpose of the Study:

  • To investigate the role of the C-terminal region of the DENV MH domain in viral replication using site-directed mutagenesis.
  • To elucidate the impact of specific amino acid substitutions on DENV particle assembly, entry, and infectivity.

Main Methods:

  • Site-directed mutagenesis was employed to introduce proline substitutions at key C-terminal residues (120, 123, 127) of the DENV MH domain.
  • Virus-like particles (VLPs) and DENV2 replicon systems were used to assess the effects of mutations on particle production and infectivity.
  • Analysis of revertant viruses was performed to understand the functional consequences of lethal mutations.

Main Results:

  • Proline substitutions at the C-terminus of the MH domain, but not the N-terminus, significantly reduced DENV VLP production across multiple DENV serotypes (DENV1, DENV2, DENV4).
  • In a DENV2 replicon system, these mutations impaired particle detection, with I123P and V127P mutations also negatively affecting viral entry.
  • When introduced into the full-length DENV2 genome, all three mutations diminished virion assembly and virus spread in cell culture.

Conclusions:

  • The C-terminus of the DENV MH domain plays a critical role in both viral assembly and entry.
  • Specific mutations (I123P, V127P) within this region are detrimental to the viral life cycle, requiring adaptive mutations for restoration.
  • These findings highlight the MH domain's C-terminus as a potential target for antiviral interventions against dengue virus.

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