Characterization of novel inhibitors of HIV-1 replication that function via alteration of viral RNA processing and rev function.
Expression of the complete HIV-1 genome depends on the appropriate processing of viral RNA. Altering the balance of viral RNA processing impairs replication of the virus. In this report, we characterize two small molecule modulators of HIV-1 RNA processing, 8-azaguanine and 2-(2-(5-nitro-2-thienyl)vinyl)quinoline (5350150), which function by distinct mechanisms to suppress viral gene expression. Although only 8-Azaguanine dramatically decreased accumulation of HIV-1 unspliced and singly spliced RNAs and altered splice site usage, both compounds blocked Gag and Env expression without affecting production of Tat (p16) and Rev regulatory proteins. Subsequent analyses suggest that these compounds affect Rev-mediated RNA transport by different mechanisms. Both compounds induced cytoplasmic accumulation of Rev, suggesting that they function, in part, by impairing Rev function. This conclusion is supported by the determination that both drugs block the nuclear export of genomic HIV-1 RNA to the cytoplasm. Testing confirmed that these compounds suppress HIV-1 expression in T cells at doses below those previously used in humans for tumour chemotherapy. Together, our observations demonstrate that small molecules can be used to inhibit HIV-1 replication by altering another avenue of viral RNA processing, offering the potential for the development of novel therapeutics for controlling this disease.
Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto M5S 1A8, Canada, Department of Molecular Genetics, University of Toronto, Toronto M5S 1A8, Canada and Department of Biochemistry, West Virginia University, Morgantown, WV 26506, USA.
Raymond W
RW
Wong
Ahalya
A
Balachandran
Matthew
M
Haaland
Peter
P
Stoilov
Alan
A
Cochrane
D019380:Anti-HIV Agents / Q000494:pharmacology*; D001375:Azaguanine / Q000494:pharmacology*; D015496:CD4-Positive T-Lymphocytes / Q000821:virology; D002460:Cell Line; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D011804:Quinolines / Q000494:pharmacology*; D012326:RNA Splicing / Q000187:drug effects*; D012367:RNA, Viral / Q000378:metabolism*; D013876:Thiophenes / Q000494:pharmacology*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects; D054321:rev Gene Products, Human Immunodeficiency Virus / Q000032:analysis / Q000037:antagonists & inhibitors*
C585478:2-(2-(5-nitro-2-thienyl)vinyl)quinoline; D019380:Anti-HIV Agents; D011804:Quinolines; D012367:RNA, Viral; D013876:Thiophenes; D015678:Viral Structural Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus; C514860:rev protein, Human Immunodeficiency Virus-1; D001375:Azaguanine