Rev is a small basic protein that is essential for HIV replication. This program outlines a comprehensive plan to answer important questions about the mechanism of Rev function, and to identify small molecule inhibitors of Rev-RRE interactions that provide the basis for developing anti-HIV therapeutics. Rev associates with a region in the viral mRNA termed the Rev Responsive Element to form an oligomeric complex that targets the mRNA for export from the nucleus, which constitutes a viable target for novel antiviral therapies. This program consists of six interrelated Projects and Core components that apply a variety of experimental approaches to study the RNA binding and nuclear export functions of Rev and to identify inhibitors of those functions. We will characterize the interactions of Rev with the RRE RNA using in vitro assays and develop screening methodologies for identification of Rev inhibitors. The disciplines that will be applied to understanding and targeting Rev include cell biology, biochemistry, structural biology, biophysics, and synthetic organic chemistry. Using this broad but well-integrated approach, we will address key remaining questions about the mechanism of Rev function, address the validity of Rev as a therapeutic target, and generate small molecule lead compounds that will stimulate drug discovery efforts involving this important but under-exploited HIV target.
| 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 |
