Binding of Tat protein to TAR region of human immunodeficiency virus type 1 blocks TAR-mediated activation of (2'-5')oligoadenylate synthetase.
The TAR sequence of the 5' leader of HIV-1 long terminal repeat-directed mRNA was found to be able to bind to and to activate double-stranded RNA-dependent (2'-5')A synthetase. Binding of TAR to the purified synthetase in vitro was abolished by addition of HIV-1 Tat protein, which binds to this sequence with a high affinity. Inhibition of TAR-mediated activation of (2'-5')A synthetase by Tat was prevented in the presence of the Zn2+ and Cd2+ chelators o-phenanthroline and penicillamine, which did not impair TAR-synthetase interaction. Transient expression assays of bacterial chloramphenicol acetyltransferase (CAT) gene in HeLa cells revealed that the levels of both CAT mRNA and CAT protein decreased after treatment of the cells with interferon, if CAT gene was linked to HIV-1 TAR segment. Cotransfection of the cells with a tat sequence containing plasmid rendered CAT gene expression insensible to the action of interferon.
Schröder
H C
HC
Institut für Physiologische Chemie, Universität, Mainz, West Germany.
Ugarković
D
D
Wenger
R
R
Reuter
P
P
Okamoto
T
T
Müller
W E
WE
D015088:2',5'-Oligoadenylate Synthetase / Q000096:biosynthesis / Q000235:genetics*; D002104:Cadmium / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004789:Enzyme Activation; D015967:Gene Expression Regulation, Viral; D015756:Genes, tat; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics*; D006801:Humans; D007372:Interferons / Q000494:pharmacology; D010396:Penicillamine / Q000494:pharmacology; D010618:Phenanthrolines / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D012091:Repetitive Sequences, Nucleic Acid; D015534:Trans-Activators / Q000378:metabolism*; D014162:Transfection; D015032:Zinc / Q000494:pharmacology
D010618:Phenanthrolines; D012333:RNA, Messenger; D015534:Trans-Activators; D002104:Cadmium; D007372:Interferons; D015500:Chloramphenicol O-Acetyltransferase; D015088:2',5'-Oligoadenylate Synthetase; D010396:Penicillamine; D015032:Zinc; C025205:1,10-phenanthroline