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Use of the Fluorescent Nucleoside Analogue Benzo[ g ]quinazoline 2′‐ O ‐Methyl‐ β ‐ D ‐ribofuranoside to Monitor the Binding of the HIV‐1 Tat Protein or of Antisense Oligonucleotides to the TAR RNA Stem‐Loop
Author(s) -
Arzumanov Andrey,
Godde Frédéric,
Moreau Serge,
Toulmé JeanJacques,
Weeds Alan,
Gait Michael J.
Publication year - 2000
Publication title -
helvetica chimica acta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.74
H-Index - 82
eISSN - 1522-2675
pISSN - 0018-019X
DOI - 10.1002/1522-2675(20000705)83:7<1424::aid-hlca1424>3.0.co;2-d
Subject(s) - chemistry , quinazoline , tar (computing) , fluorescence , oligonucleotide , rna , nucleoside , activator (genetics) , transcription (linguistics) , peptide , human immunodeficiency virus (hiv) , stereochemistry , biochemistry , gene , programming language , physics , quantum mechanics , computer science , medicine , linguistics , philosophy , family medicine
The Tat protein is an essential trans ‐activator of HIV gene expression. It interacts with its RNA recognition sequence, the trans‐ activation responsive region TAR, as well as cellular factors. These interactions are potential targets for drug discovery against HIV infection. We have developed a new and sensitive assay for the measurement of Tat binding to TAR in solution under equilibrium conditions based on the change of fluorescence of the base analogue benzo[ g ]quinazoline‐2,4(1 H ,3 H )‐dione (BgQ) incorporated into the chemically synthesized model TAR stem‐loop 2 to which was added Tat‐[37‐72] peptide ( 3 ). The results show that Tat‐TAR binding strength is 2 – 3‐fold stronger than has previously been determined by mobility‐shift analysis. Changes of fluorescence were used also to measure the binding of antisense 2′‐ O ‐methyloligonucleotides to TAR 2 .