Immunolocalization of Picornavirus RNA in infected cells with antibodies to Tyr-pUp, the covalent linkage unit between VPg and RNA

Elena S. Gavryushina, Sofia A. Bryantseva, Elena S. Nadezhdina, Timofei S. Zatsepin, Ilya Yu Toropygin, Angela Pickl-Herk, Dieter Blaas, Yuri F. Drygin

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

The genomic RNA of picornaviruses is attached to a small protein (VPg) via a covalent bond between a tyrosine and a 5'-terminal uridine phosphate. The same structure is present in potyvirus and calicivirus families. VPgs play a key role in initiation of viral replication by acting as primers for RNA synthesis. The model compound [N(Ac),CO(NHMe)]Tyr-(5'P→O)Up-O-(CH2)6NH2 (mCLU), mimicking this 'covalent linkage unit' (CLU) and containing Tyr-pUp was synthesized in solution following the phosphoramidite scheme and used to raise antibodies for studying picornavirus infection. The antibodies recognized CLU-containing mengovirus RNA and showed minimal cross-reactivity with RNAs lacking CLU. Immunofluorescence staining of cells infected with a human rhinovirus demonstrated co-localization of the signals from anti-mCLU and from anti-VPg antibodies. Efficient synthesis of mCLU and anti-mCLU antibodies might be of great utility for investigating viral replication and identifying yet unknown viral and cellular CLU-containing RNA-protein complexes.

Original languageEnglish
Pages (from-to)206-211
Number of pages6
JournalJournal of Virological Methods
Volume171
Issue number1
DOIs
Publication statusPublished - Jan 2011
Externally publishedYes

Keywords

  • Covalent linkage unit
  • Dot-blot immunogold assay
  • Human rhinovirus
  • Immunofluorescence microscopy
  • Picornavirus
  • VPg-RNA

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