NMR solution structure of poliovirus uridylyated peptide linked to the genome (VPgpU)

Catherine H. Schein, Numan Oezguen, Gerbrand J. Van Der Heden Van Noort, Dmitri V. Filippov, Aniko Pauld, Eric Kumar, Werner Braun

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Picornaviruses have a 22-24 amino acid peptide, VPg, bound covalently at the 5' end of their RNA, that is essential for replication. VPgs are uridylylated at a conserved tyrosine to form VPgpU, the primer of RNA synthesis by the viral polymerase. This first complete structure for any uridylylated VPg, of poliovirus type 1 (PV1)-VPgpU, shows that conserved amino acids in VPg stabilize the bound UMP, with the uridine atoms involved in base pairing and chain elongation projected outward. Comparing this structure to PV1-VPg and partial structures of VPg/VPgpU from other picornaviruses suggests that enteroviral polymerases require a more stable VPg structure than does the distantly related aphthovirus, foot and mouth disease virus (FMDV). The glutamine residue at the C-terminus of PV1-VPgpU lies in back of the uridine base and may stabilize its position during chain elongation and/or contribute to base specificity. Under in vivo-like conditions with the authentic cre(2C) hairpin RNA and Mg 2+, 5-methylUTP cannot compete with UTP for VPg uridylyation in an in vitro uridylyation assay, but both nucleotides are equally incorporated by PV1-polymerase with Mn 2+ and a poly-A RNA template. This indicates the 5 position is recognized under in vivo conditions. The compact VPgpU structure docks within the active site cavity of the PV-polymerase, close to the position seen for the fragment of FMDV-VPgpU with its polymerase. This structure could aid in design of novel enterovirus inhibitors, and stabilization upon uridylylation may also be pertinent for post-translational uridylylation reactions that underlie other biological processes.

Original languageEnglish (US)
Pages (from-to)1441-1448
Number of pages8
JournalPeptides
Volume31
Issue number8
DOIs
StatePublished - Aug 2010

Fingerprint

Poliovirus
Uridine
Viruses
Elongation
Genes
Nuclear magnetic resonance
Genome
RNA
Uridine Monophosphate
Picornaviridae
Amino Acids
Foot-and-Mouth Disease Virus
Peptides
Uridine Triphosphate
Docks
Glutamine
Tyrosine
Aphthovirus
Assays
Nucleotides

Keywords

  • Antiviral compounds
  • Disordered structures
  • Enteroviruses
  • Flexibility
  • Picornaviruses
  • Polymerase priming mechanism
  • Post-translational modification
  • RNA editing
  • Uridylylation

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Physiology
  • Cellular and Molecular Neuroscience

Cite this

Schein, C. H., Oezguen, N., Van Der Heden Van Noort, G. J., Filippov, D. V., Pauld, A., Kumar, E., & Braun, W. (2010). NMR solution structure of poliovirus uridylyated peptide linked to the genome (VPgpU). Peptides, 31(8), 1441-1448. https://doi.org/10.1016/j.peptides.2010.04.021

NMR solution structure of poliovirus uridylyated peptide linked to the genome (VPgpU). / Schein, Catherine H.; Oezguen, Numan; Van Der Heden Van Noort, Gerbrand J.; Filippov, Dmitri V.; Pauld, Aniko; Kumar, Eric; Braun, Werner.

In: Peptides, Vol. 31, No. 8, 08.2010, p. 1441-1448.

Research output: Contribution to journalArticle

Schein, CH, Oezguen, N, Van Der Heden Van Noort, GJ, Filippov, DV, Pauld, A, Kumar, E & Braun, W 2010, 'NMR solution structure of poliovirus uridylyated peptide linked to the genome (VPgpU)', Peptides, vol. 31, no. 8, pp. 1441-1448. https://doi.org/10.1016/j.peptides.2010.04.021
Schein CH, Oezguen N, Van Der Heden Van Noort GJ, Filippov DV, Pauld A, Kumar E et al. NMR solution structure of poliovirus uridylyated peptide linked to the genome (VPgpU). Peptides. 2010 Aug;31(8):1441-1448. https://doi.org/10.1016/j.peptides.2010.04.021
Schein, Catherine H. ; Oezguen, Numan ; Van Der Heden Van Noort, Gerbrand J. ; Filippov, Dmitri V. ; Pauld, Aniko ; Kumar, Eric ; Braun, Werner. / NMR solution structure of poliovirus uridylyated peptide linked to the genome (VPgpU). In: Peptides. 2010 ; Vol. 31, No. 8. pp. 1441-1448.
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