Human immunodeficiency virus tat-activated expression of poliovirus protein 2A inhibits mRNA translation.
To study the effect of poliovirus protein 2A on cellular RNA translation, the tat control system of human immunodeficiency virus (HIV) was used. Protein 2A was expressed from a plasmid construct (pHIV/2A) incorporating the HIV long terminal repeat. Protein synthesis was measured by using chloramphenicol acetyltransferase as a reporter gene driven by the Rous sarcoma virus long terminal repeat. When HIV/2A was cotransfected with the reporter, addition of a tat-producing plasmid caused at least a 50-fold drop in chloramphenicol acetyltransferase synthesis. A HeLa cell line carrying HIV/2A was established. In it, tat expression caused more than a 10-fold drop in chloramphenicol acetyltransferase synthesis from the reporter plasmid. Furthermore, 2A induction by tat caused cleavage of the cellular translation factor P220, a part of eukaryotic translation initiation factor 4F. Thus protein 2A can, by itself, carry out the inhibition of cellular protein synthesis characteristic of a poliovirus infection. Also, the HIV tat activation provides a very effective method to control gene expression in mammalian cells.
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Whitehead Institute for Biomedical Research, MA 02142.
Baltimore
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D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000235:genetics*; D039562:Eukaryotic Initiation Factor-4F; D005786:Gene Expression Regulation; D005814:Genes, Viral; D006678:HIV / Q000235:genetics*; D006367:HeLa Cells / Q000378:metabolism; D008970:Molecular Weight; D010448:Peptide Initiation Factors / Q000096:biosynthesis / Q000235:genetics; D010957:Plasmids; D017955:Poliovirus / Q000235:genetics*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D015183:Restriction Mapping; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins
D039562:Eukaryotic Initiation Factor-4F; D010448:Peptide Initiation Factors; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D015500:Chloramphenicol O-Acetyltransferase; D003546:Cysteine Endopeptidases; C057400:picornain 2A, Picornavirus