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Reverse Genetics Mediated Recovery of Infectious Murine Norovirus
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Norovirus VPg Binds RNA through a Conserved N-Terminal K/R Basic Patch.

Alice M McSweeney1, Vivienne L Young1, Vernon K Ward1

  • 1Department of Microbiology & Immunology, School of Biomedical Sciences, University of Otago, P.O. Box 56, Dunedin 9054, New Zealand.

Viruses
|July 2, 2021
PubMed
Summary
This summary is machine-generated.

Murine norovirus (MNV) genome-linked protein (VPg) binds RNA via a basic patch in its N-terminal region. This RNA binding is crucial for MNV VPg to trigger cell cycle arrest.

Keywords:
RNA bindingVPgcalicivirusnorovirusspidroin

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Area of Science:

  • Virology
  • Molecular Biology
  • Structural Biology

Background:

  • The viral protein genome-linked (VPg) is essential for norovirus replication.
  • Norovirus VPg proteins possess RNA-binding capabilities, particularly within their N-terminal regions.

Purpose of the Study:

  • To investigate the RNA-binding properties of murine norovirus (MNV) VPg.
  • To identify the specific amino acid residues responsible for RNA interaction.
  • To determine the role of RNA binding in MNV VPg-induced cell cycle arrest.

Main Methods:

  • Expression of MNV and human norovirus (HuNV) VPg proteins fused to a spidroin tag in insect cells.
  • Electrophoretic mobility shift assays (EMSA) using pentaprobe RNA sequences to assess RNA binding.
  • Site-directed mutagenesis of basic residues (lysine/arginine) in the MNV VPg N-terminus.
  • Analysis of MNV VPg's effect on cell cycle progression (G0/G1 arrest).

Main Results:

  • MNV VPg and HuNV VPg proteins exhibit non-specific RNA binding.
  • A basic patch comprising lysine/arginine residues within the first 12 amino acids of MNV VPg is critical for RNA binding.
  • Mutation of this basic patch abolishes RNA binding.
  • The RNA-binding deficient MNV VPg mutant fails to induce G0/G1 cell cycle arrest.

Conclusions:

  • Norovirus VPg proteins utilize a disordered N-terminal basic patch for non-specific RNA binding.
  • This RNA interaction is a key mechanism by which MNV VPg influences host cell cycle progression.
  • Understanding VPg-RNA interactions offers potential targets for antiviral strategies against noroviruses.