This research project is part of a comprehensive program-Targeting the RNA binding and nuclear export of HIV Rev-aimed at characterizing HIV Rev function and identifying new approaches for inhibiting the replication of HIV. The objective of this research component is to synthesize compounds that will be screened for their capacity to inhibit the binding of Rev to RRE RNA and Rev-dependent mRNA export. This proposal describes strategies for synthesizing analogues of the naturally occurring HIV Rev/RRE RNA inhibitors harziphilone and fleephilone as well as a wide variety of drug-like molecules with RNA-binding potential from a virtual library developed by Williamson in project #1. Syntheses of 13C-labeled variants of harziphilone and fleephilone will facilitate NMR studies to elucidate the structures of the complexes that these natural products make with the RRE of viral mRNA or with a Rev-RRE complex. The structural information yielded by these studies will guide the design and synthesis of analogues of harziphilone and fleephilone in an effort to understand the molecular features critical for potency and selectivity. In addition, both novel and precedented multi-component coupling reactions will be applied to syntheses of manifold small molecules that have drug-like characteristics. These compounds will be evaluated by Williamson project #1, Millar project #2, and Gerace project #4 for their utility as inhibitors of HIV Rev function. This project will contribute a collection of novel molecules possessing RNA-binding and drug-like features to a multi-disciplinary collaboration that will explore the therapeutic potential of disrupting the binding interaction between the HIV-1 Rev protein and the RRE of viral mRNA. Ultimately, this research could furnish new lead compounds for the development of new anti-viral agents against HIV-infection.
| Marenchino, Marco; Armbruster, David W; Hennig, Mirko (2009) Rapid and efficient purification of RNA-binding proteins: application to HIV-1 Rev. Protein Expr Purif 63:112-9 |
| Pond, Stephanie J K; Ridgeway, William K; Robertson, Rae et al. (2009) HIV-1 Rev protein assembles on viral RNA one molecule at a time. Proc Natl Acad Sci U S A 106:1404-8 |
| Carlomagno, Teresa; Amata, Irene; Williamson, James R et al. (2008) NMR assignments of HIV-2 TAR RNA. Biomol NMR Assign 2:167-9 |
| Pljevaljcic, Goran; Millar, David P (2008) Single-molecule fluorescence methods for the analysis of RNA folding and ribonucleoprotein assembly. Methods Enzymol 450:233-52 |
| Edgcomb, Stephen P; Aschrafi, Angelique; Kompfner, Elizabeth et al. (2008) Protein structure and oligomerization are important for the formation of export-competent HIV-1 Rev-RRE complexes. Protein Sci 17:420-30 |
| Scott, Lincoln G; Hennig, Mirko (2008) RNA structure determination by NMR. Methods Mol Biol 452:29-61 |
| Hennig, Mirko; Scott, Lincoln G; Sperling, Edit et al. (2007) Synthesis of 5-fluoropyrimidine nucleotides as sensitive NMR probes of RNA structure. J Am Chem Soc 129:14911-21 |
| Hennig, Mirko; Munzarova, Marketa L; Bermel, Wolfgang et al. (2006) Measurement of long-range 1H-19F scalar coupling constants and their glycosidic torsion dependence in 5-fluoropyrimidine-substituted RNA. J Am Chem Soc 128:5851-8 |
