Inhibition of HIV-1 Rev-RRE interaction by diphenylfuran derivatives.
The interactions between RNA structures, such as RRE in the HIV-1 genome, and proteins, such as Rev of HIV-1, are essential for efficient viral replication. Compounds that bind specifically to such RNAs and disrupt their protein complexes offer a novel mechanism for inhibition of replication of the virus. As a step in this approach, we have designed and characterized a series of synthetic diphenylfuran cations that selectively inhibit Rev binding to RRE. Fluorescence titrations and gel band-shift results indicate that the diphenylfurans bind to RRE and inhibit Rev complex formation in a structure-dependent manner. The derivative with the greatest affinity for RRE has an association constant of greater than 10(7) M-1 and inhibits formation of the Rev--RRE complex at concentrations below 1 microM. It binds to RRE considerably more strongly than it binds to simple RNA duplexes. Spectral changes and energy transfer results on complex formation suggest that the compound has a nonclassical intercalation binding mode. CD studies with modified RRE hairpins indicate that the active diphenylfurans bind at the structured internal loop of RRE and cause a conformational change. The most active diphenylfurans are tetracations that appear to bind to RRE by a threading intercalation mode and cause a conformational change in the RNA that is essential for inhibition of Rev complex formation with RRE.
Ratmeyer
L
L
Department of Chemistry, Georgia State University, Atlanta 30303, USA.
Zapp
M L
ML
Green
M R
MR
Vinayak
R
R
Kumar
A
A
Boykin
D W
DW
Wilson
W D
WD
D000578:Amidines / Q000494:pharmacology; D019380:Anti-HIV Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D001550:Benzamidines; D002942:Circular Dichroism; D005663:Furans / Q000378:metabolism / Q000494:pharmacology*; D015694:Gene Products, rev / Q000378:metabolism*; D015752:Genes, env; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics; D007364:Intercalating Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D007700:Kinetics; D015394:Molecular Structure; D009690:Nucleic Acid Conformation / Q000187:drug effects; D009691:Nucleic Acid Denaturation; D012367:RNA, Viral / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D054321:rev Gene Products, Human Immunodeficiency Virus
D000578:Amidines; D019380:Anti-HIV Agents; D001550:Benzamidines; D005663:Furans; D015694:Gene Products, rev; D007364:Intercalating Agents; D012367:RNA, Viral; D054321:rev Gene Products, Human Immunodeficiency Virus; C033286:furamidine