Selection and characterization of small molecules that bind the HIV-1 frameshift site RNA.
HIV-1 requires a -1 translational frameshift to properly synthesize the viral enzymes required for replication. The frameshift mechanism is dependent upon two RNA elements, a seven-nucleotide slippery sequence (UUUUUUA) and a downstream RNA structure. Frameshifting occurs with a frequency of approximately 5%, and increasing or decreasing this frequency may result in a decrease in viral replication. Here, we report the results of a high-throughput screen designed to find small molecules that bind to the HIV-1 frameshift site RNA. Out of 34,500 compounds screened, 202 were identified as positive hits. We show that one of these compounds, doxorubicin, binds the HIV-1 RNA with low micromolar affinity (K(d) = 2.8 microM). This binding was confirmed and localized to the RNA using NMR. Further analysis revealed that this compound increased the RNA stability by approximately 5 degrees C and decreased translational frameshifting by 28% (+/-14%), as measured in vitro.
Marcheschi
Ryan J
RJ
Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA.
Mouzakis
Kathryn D
KD
Butcher
Samuel E
SE
D001483:Base Sequence; D001665:Binding Sites; D004317:Doxorubicin / Q000494:pharmacology; D018965:Frameshifting, Ribosomal / Q000187:drug effects*; D015658:HIV Infections / Q000188:drug therapy*; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology*
D012367:RNA, Viral; D054852:Small Molecule Libraries; D004317:Doxorubicin