Sequence specificity for uridylylation of the viral peptide linked to the genome (VPg) of enteroviruses

Related Experiment Video

Updated: Apr 10, 2026

Analysis of Group IV Viral SSHHPS Using In Vitro and In Silico Methods
10:40

Analysis of Group IV Viral SSHHPS Using In Vitro and In Silico Methods

Published on: December 21, 2019

26.6K

Catherine H Schein1, Mengyi Ye2, Aniko V Paul3

  • 1Foundation for Applied Molecular Evolution (FfAME), 13709 Progress Blvd., Alachua, FL 32616, United States.

Virology
|June 16, 2015
PubMed

Insights

Enteroviruses uridylylate a viral protein (VPg) to initiate RNA replication. Diverse enterovirus polymerases (3Dpol) can uridylylate various VPg peptides, suggesting conserved mechanisms for this crucial step.

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Enteroviruses (EV) initiate replication by uridylylating a protein primer, VPg.
  • The uridylylated VPg (VPgpUpU) functions as the primer for viral RNA synthesis.

Purpose of the Study:

  • To compare the uridylylation abilities of four different enterovirus polymerases (3Dpol) from species A-C.
  • To investigate the substrate specificity of EV 3Dpol for VPg peptides with significant sequence variation.

Main Methods:

  • In vitro uridylylation assays using four EV 3Dpol enzymes and five VPg variants.
  • Utilized polyA RNA as a template for uridylylation.
  • Compared VPg-binding sites through co-crystal structure analysis.

Main Results:

  • All tested 3Dpol enzymes uridylylated all tested VPg peptides using polyA RNA as a template.
  • Each 3Dpol demonstrated specificity for its cognate VPg.
  • A consensus VPg, reflecting conserved physicochemical properties, was efficiently uridylylated by all 3Dpol, indicating conserved requirements for uridylylation.
  • Analysis of conserved polymerase residues impacting uridylylation but not elongation.

Conclusions:

  • The uridylylation step in enterovirus replication involves conserved residues and enzymatic mechanisms across diverse EV species.
  • The reaction likely involves a second 3Dpol molecule due to differing VPg-binding sites observed in co-crystal structures.
Keywords:
Coxsackie virusEV-71Metal ion dependent phosphotransferNucleotide transfer reactionPCP-consensus sequencePeptide primingPoliovirusRNA polymerase

More Related Videos

Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting08:40

Production of Pseudotyped Particles to Study Highly Pathogenic Coronaviruses in a Biosafety Level 2 Setting

Published on: March 1, 2019

59.8K
Surface Functionalization of Hepatitis E Virus Nanoparticles Using Chemical Conjugation Methods09:12

Surface Functionalization of Hepatitis E Virus Nanoparticles Using Chemical Conjugation Methods

Published on: May 11, 2018

7.4K

Related Concept Videos

Leaky Scanning02:28

Leaky Scanning

During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA.  Marilyn Kozak discovered that the sequence RCCAUGG (where R...
5.9K
Size and Structure of Viral Genomes01:26

Size and Structure of Viral Genomes

Viral genomes exhibit remarkable diversity in size, structure, and composition, influencing their replication strategies and interactions with host cells. These genomes consist of either DNA or RNA and may be linear or circular. Additionally, they can be single-stranded or double-stranded, with each configuration affecting how the virus propagates within a host. RNA viruses, for instance, generally have smaller genomes than DNA viruses, a factor that contributes to their high mutation rates and...
1.2K
Viruses with RNA Genomes01:29

Viruses with RNA Genomes

RNA viruses are categorized into positive-strand, negative-strand, or double-stranded groups based on their genomic structure and replication mechanisms. This classification dictates how they exploit host cellular machinery for protein synthesis and replication. Some RNA viruses also utilize reverse transcription as part of their life cycle, further diversifying their replication strategies.Positive-Strand RNA VirusesPositive-strand RNA viruses have genomes that function directly as messenger...
1.4K