Document Detail

Immunolocalization of Picornavirus RNA in infected cells with antibodies to Tyr-pUp, the covalent linkage unit between VPg and RNA.
MedLine Citation:
PMID:  21056058     Owner:  NLM     Status:  MEDLINE    
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-(CH(2))(6)NH(2) (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.
Elena S Gavryushina; Sofia A Bryantseva; Elena S Nadezhdina; Timofei S Zatsepin; Ilya Yu Toropygin; Angela Pickl-Herk; Dieter Blaas; Yuri F Drygin
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-11-04
Journal Detail:
Title:  Journal of virological methods     Volume:  171     ISSN:  1879-0984     ISO Abbreviation:  J. Virol. Methods     Publication Date:  2011 Jan 
Date Detail:
Created Date:  2010-12-28     Completed Date:  2011-04-04     Revised Date:  2011-10-04    
Medline Journal Info:
Nlm Unique ID:  8005839     Medline TA:  J Virol Methods     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  206-11     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 Elsevier B.V. All rights reserved.
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, Russia.
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MeSH Terms
Antibodies, Viral / diagnostic use*,  isolation & purification
Hela Cells
Microscopy, Fluorescence / methods
Oligoribonucleotides / chemical synthesis*,  immunology*
Picornaviridae / chemistry,  growth & development*
RNA, Viral / analysis*
Virology / methods*
Grant Support
P 18693-B09//Austrian Science Fund FWF
Reg. No./Substance:
0/Antibodies, Viral; 0/Oligoribonucleotides; 0/RNA, Viral; 0/VPg-pUpU

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