Document Detail


A stable HeLa cell line that inducibly expresses poliovirus 2A(pro): effects on cellular and viral gene expression.
MedLine Citation:
PMID:  10666269     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A HeLa cell clone (2A7d) that inducibly expresses the gene for poliovirus protease 2A (2A(pro)) under the control of tetracycline has been obtained. Synthesis of 2A(pro) induces severe morphological changes in 2A7d cells. One day after tetracycline removal, cells round up and a few hours later die. Poliovirus 2A(pro) cleaves both forms of initiation factor eIF4G, causing extensive inhibition of capped-mRNA translation a few hours after protease induction. Methoxysuccinyl-Ala-Ala-Pro-Val-chloromethylketone, a selective inhibitor of 2A(pro), prevents both eIF4G cleavage and inhibition of translation but not cellular death. Expression of 2A(pro) still allows both the replication of poliovirus and the translation of mRNAs containing a picornavirus leader sequence, while vaccinia virus replication is drastically inhibited. Translation of transfected capped mRNA is blocked in 2A7d-On cells, while luciferase synthesis from a mRNA bearing a picornavirus internal ribosome entry site (IRES) sequence is enhanced by the presence of 2A(pro). Moreover, synthesis of 2A(pro) in 2A7d cells complements the translational defect of a poliovirus 2A(pro)-defective variant. These results show that poliovirus 2A(pro) expression mimics some phenotypical characteristics of poliovirus-infected cells, such as cell rounding, inhibition of protein synthesis and enhancement of IRES-driven translation. This cell line constitutes a useful tool to further analyze 2A(pro) functions, to complement poliovirus 2A(pro) mutants, and to test antiviral compounds.
Authors:
A Barco; E Feduchi; L Carrasco
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of virology     Volume:  74     ISSN:  0022-538X     ISO Abbreviation:  J. Virol.     Publication Date:  2000 Mar 
Date Detail:
Created Date:  2000-11-03     Completed Date:  2000-11-03     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  0113724     Medline TA:  J Virol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  2383-92     Citation Subset:  IM    
Affiliation:
Centro de Biología Molecular, Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
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MeSH Terms
Descriptor/Qualifier:
Cysteine Endopeptidases / biosynthesis,  genetics,  metabolism*
Eukaryotic Initiation Factor-4G
Gene Expression Regulation, Viral*
Hela Cells / metabolism,  virology*
Humans
Immunoblotting
Peptide Initiation Factors / analysis,  metabolism
Poliovirus
Promoter Regions, Genetic / drug effects
Protease Inhibitors / pharmacology
Protein Biosynthesis / drug effects
Protein Synthesis Inhibitors / metabolism*
RNA, Messenger / genetics
Tetracycline / pharmacology
Transfection
Viral Proteins*
Virus Replication
Chemical
Reg. No./Substance:
0/Eukaryotic Initiation Factor-4G; 0/Peptide Initiation Factors; 0/Protease Inhibitors; 0/Protein Synthesis Inhibitors; 0/RNA, Messenger; 0/Viral Proteins; 60-54-8/Tetracycline; EC 3.4.22.-/Cysteine Endopeptidases; EC 3.4.22.29/picornain 2A, Picornavirus
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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