Elucidation and characterization of oligonucleotide-accessible sites on HIV-2 leader region RNA.
The retroviruses, including the human pathogens HIV-1 and HIV-2, are diploid inasmuch as they encapsidate two copies of their RNA genome. Prior to or during encapsidation, two copies of full-length genomic RNA recognize and stably bind each other in a process called dimerization. RNA structures within the viral genome promote dimerization in both HIV-1 and HIV-2 and are located in the 5'-untranslated leader region. Inhibition of dimerization by mutation of these RNA signals has been demonstrated to drastically reduce viral infectivity and replication kinetics and, thus, represents a potential target for antiretroviral therapy. In this study, we identified sites in HIV-2 leader region RNA that are functionally accessible to hybridization with oligonucleotides (ODNs) by reverse transcription with random ODN libraries (RT-ROL). We then tested specific ODNs directed against these regions for their efficacy in inhibiting RNA dimerization in vitro. We determined that of several hybridization-competent ODNs, only two were very effective in inhibiting RNA dimerization. Both of these ODNs were complementary to viral RNA at the primer binding site (PBS). These results identify regions with high accessibility to ODN binding on HIV-2 RNA and help to map the region(s) essential for dimerization within the viral RNA.
Deer
Emily L
EL
Division of Biological Sciences, Biochemistry/Microbiology Graduate Program, The University of Montana, Missoula, MT 59812, USA.
Douk
Boramee
B
Lanchy
Jean-Marc
JM
Lodmell
J Stephen
JS
D020121:5' Untranslated Regions; D015498:HIV-2 / Q000235:genetics*; D006801:Humans; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D012313:RNA / Q000378:metabolism*; D016914:Ribonuclease H / Q000378:metabolism; D017423:Sequence Analysis, RNA
D020121:5' Untranslated Regions; D009838:Oligodeoxyribonucleotides; D012313:RNA; D016914:Ribonuclease H