Immunolocalization of Picornavirus RNA in infected cells with antibodies to Tyr-pUp, the covalent linkage unit between VPg and RNA.
J Virol Methods
; 171(1): 206-11, 2011 Jan.
Article
in En
| MEDLINE
| ID: mdl-21056058
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'PâO)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.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Oligoribonucleotides
/
Picornaviridae
/
Virology
/
RNA, Viral
/
Antibodies, Viral
Limits:
Animals
/
Humans
Language:
En
Journal:
J Virol Methods
Year:
2011
Document type:
Article
Affiliation country:
RUSSIA