Small molecules that selectively block RNA binding of HIV-1 rev protein inhibit rev function and viral production

Inhibition of protein synthesis by aminoglycoside???arginine conjugates

“…RNA contains complex and sophisticated higher order structures that are essential for recognition by other macromolecules and/or required for catalytic processes+ As such, it offers an interesting target for small molecule ligands and, indeed, the ability of RNA to interact with small molecules has long been recognized+ Antibiotics are a chemically and structurally diverse collection of molecules, with some classes capable of interacting with rRNA to exert profound effects on the translation process+ Functional insights from structural studies of compounds bound to ribosomal subunits have revealed that rRNA/small molecule ligand recognition is based on a combination of shape recognition, electrostatic, and hydrogen-bonding interactions Pioletti et al+, 2001;Schlunzen et al+, 2001)+ Additionally, RNA SELEX (Systematic Evolution of Ligands by Exponential Enrichment) has enabled the identification of minimal nucleic acid recognition motifs for ligand binding, demonstrating that RNA three-dimensional structures can form a large number of highly specific ligand-binding sites (Gold et al+, 1995)+ The ribosome is the target for many important antibacterial agents; these compounds interfere with essential steps of protein synthesis (Pestka, 1977;Gale et al+, 1981;Noller, 1991)+ Among these, 2-deoxystreptamine aminoglycosides (small polycationic compounds possessing linked ring systems consisting of aminosugars and an aminocyclitol) cause codon misreading by interfering with the decoding process + These compounds have found clinical use as antibacterial agents due to their ability to specifically bind bacterial ribosomes (Gale et al+, 1981) and are thought to exert their effects by increasing the error rate of the ribosome + Eukary-otic cytoplasmic ribosomes are relatively insensitive to 2-deoxystreptamine aminoglycosides (Kurtz, 1974;Palmer & Wilhelm, 1978;Wilhelm et al+, 1978aWilhelm et al+, , 1978b, and it has been suggested that the sensitivity of a ribosomal system to antibiotics is determined by the sequence of its rRNA (Sor & Fukuhara, 1984;Beckers et al+, 1995)+ Indeed, the nephrotoxicity and ototoxicity associated with use of aminoglycosides in the clinical setting may be linked to the susceptibility of mitochondrial ribosomes to these compounds (Bottger et al+, 2001)+ Aminoglycosides are also capable of binding to and affecting the activity of a large number of other RNAs, including the HIV TAR element (Mei et al+, 1997), HIV Rev-responsive element (Zapp et al+, 1993), hammerhead ribozymes (Stage et al+, 1995;Jenne et al+, 2001), hairpin ribozymes (Earnshaw & Gait, 1998), ribonuclease P RNA …”

Aminoglycoside binding to the hammerhead ribozyme: a general model for the interaction of cationic antibiotics with RNA 1 1Edited by J. Karn

“…The neomycins bind to functional sites in the 16 S ribosomal RNA and cause miscoding and translocation arrest of the ribosome (Moazed & Noller, 1987). Neomycin B inhibits also human immunode®ciency virus (HIV) replication in vivo by selectively blocking the binding of the Rev protein to its viral RNA target (Zapp et al, 1993). Other RNA-catalysed processes found to be inhibited by aminoglycoside antibiotics include the self-splicing of group I introns (von Ahsen et al, 1991), the cleavage reactions of the hammerhead ribozyme and the hepatitis delta virus (HDV) ribozyme (Rogers et al, 1996).…”

A reporter based single step assay for evaluation of inhibitors targeting HIV-1 Rev–RRE interaction

“…The classical approach towards drug discovery has been the screening of a vast number of compounds/drugs and this approach also has been utilized to discover lead compounds capable of inhibiting Rev-RRE system. A number of small-molecule compounds, aminoglycoside antibiotics such as neomycin, RRE decoys, transdominant-negative version of the Rev protein and diphenylfuran cations have been screened for inhibiton of Rev-RRE interaction [9,11,14,22,23,28]. Most of the screening assays described so far are based on transfections, require a lot of experimental manipulations and are time consuming.…”

“…Proteins were separated on a 15 % SDS-PAGE gel, transferred to PVDF membrane (Immobilon-P; Millipore, USA). The membranes were blocked (5 % non fat dry milk in Tris-buffered saline with 0.1 % Tween 20) for 1 h followed by overnight incubation at 4°C with HIV-1 Rev antibody [8,28]. Membrane was washed and incubated with HRPO conjugated secondary Fig.…”