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T135I substitution in the nonstructural protein 2C enhances foot-and-mouth disease virus replication

Tiangang Yuan1, Haiwei Wang2, Chen Li1

  • 1Division of Livestock Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 678 Haping Road, Xiangfang District, Harbin, 150069, People's Republic of China.

Virus Genes
|June 22, 2017
PubMed

Insights

A 60-amino acid deletion in foot-and-mouth disease virus (FMDV) nonstructural protein 3A impaired viral replication. However, a substitution in the 2C protein (T135I) restored replication and enhanced viral RNA synthesis.

Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Foot-and-mouth disease virus (FMDV) nonstructural protein 3A is crucial for viral replication, virulence, and host range.
  • Deletions in the 3A protein's C-terminal half can attenuate FMDV, affecting replication in different cell types.

Purpose of the Study:

  • To investigate the impact of a significant 3A protein deletion on FMDV replication.
  • To identify viral genetic changes that compensate for the 3A deletion and restore replication.

Main Methods:

  • Construction and rescue of a 60-amino acid 3A deletion FMDV mutant (O/YS/CHA/05-Δ3A).
  • Passaging the mutant virus in cell culture and analyzing its replication kinetics.
  • Sequencing the passaged virus to identify genetic mutations.
  • Site-directed mutagenesis to introduce specific mutations (VP1 P153L, 2C T135I) and assess their effect on viral replication.

Main Results:

  • The 60-amino acid deletion in 3A significantly impaired FMDV replication and plaque formation.
  • After passaging, the virus regained wild-type replication levels, acquiring mutations.
  • The 2C T135I substitution, but not VP1 P153L, restored and enhanced viral replication.
  • The 2C T135I substitution enhanced FMDV replication independently of the 3A deletion and promoted viral RNA synthesis.

Conclusions:

  • The 60-amino acid deletion in FMDV 3A protein attenuates the virus.
  • The nonstructural protein 2C T135I substitution is a key factor in restoring and enhancing FMDV replication.
  • The 2C T135I substitution enhances viral RNA synthesis, contributing to increased FMDV replication capacity.
Keywords:
Foot-and-mouth disease virusMutations and growth kineticsRNA synthesisVirus replication

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