We propose to identify and develop small molecule inhibitors that target two previously neglected stages of the HIV life cycle: virus budding and capsid assembly. In the first aim, we will focus on developing inhibitors of the essential interaction between the cellular protein Tsgl01 and the p6 domain of the HIV-1 Gag protein. In preliminary studies, we have discovered that human Tsgl01 performs an essential role in virus budding by binding to the p6 domain of the HIV Gag protein and recruiting other cellular factors in the Vps pathway that facilitate virus release. We have also determined the solution structure of the p6-binding UEV domain of Tsgl 01 . Our collaborators at Myriad Pharmaceuticals have discovered lead compounds that inhibit the Tsgl01 UEV/p6 interaction. We will now determine the structure of Tsgl01 UEV in complex with its p6 binding site and in complex with Myriad's inhibitors as part of a comprehensive program in structure-assisted development of inhibitors of HIV budding. In the second aim, we will focus on discovering small molecule inhibitors of HIV capsid assembly. In principle, viral capsid assembly represents an attractive target for therapeutic intervention, but the lack of suitable assays for discovering lead compounds has been a major impediment in this research. In preliminary studies, we have determined structures of the HIV CA protein in two different conformational states, characterized the higher order assemblies formed by recombinant CA proteins, and overcome a number of hurdles in inhibitor assay development. We now propose to pursue the discovery of HIV capsid assembly inhibitors by adapting our assays to run on high-throughput platforms, screening chemical libraries for small molecule inhibitors, and characterizing the biochemistry and structural biology of the lead compounds that emerge from our screens.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Program Projects (P01)
Project #
1P01GM066521-01
Application #
6553717
Study Section
Special Emphasis Panel (ZRG1)
Project Start
2002-06-15
Project End
2007-06-14
Budget Start
Budget End
Support Year
1
Fiscal Year
2002
Total Cost
Indirect Cost
Name
University of Utah
Department
Type
DUNS #
City
Salt Lake City
State
UT
Country
United States
Zip Code
84112
Bayro, Marvin J; Ganser-Pornillos, Barbie K; Zadrozny, Kaneil K et al. (2016) Helical Conformation in the CA-SP1 Junction of the Immature HIV-1 Lattice Determined from Solid-State NMR of Virus-like Particles. J Am Chem Soc 138:12029-32
Votteler, Jörg; Sundquist, Wesley I (2013) Virus budding and the ESCRT pathway. Cell Host Microbe 14:232-41
Gan, Lu; Jensen, Grant J (2012) Electron tomography of cells. Q Rev Biophys 45:27-56
Guerrero-Ferreira, Ricardo C; Viollier, Patrick H; Ely, Bert et al. (2011) Alternative mechanism for bacteriophage adsorption to the motile bacterium Caulobacter crescentus. Proc Natl Acad Sci U S A 108:9963-8
Dobro, Megan J; Melanson, Linda A; Jensen, Grant J et al. (2010) Plunge freezing for electron cryomicroscopy. Methods Enzymol 481:63-82
Briegel, Ariane; Chen, Songye; Koster, Abraham J et al. (2010) Correlated light and electron cryo-microscopy. Methods Enzymol 481:317-41
Chen, Songye; McDowall, Alasdair; Dobro, Megan J et al. (2010) Electron cryotomography of bacterial cells. J Vis Exp :
Rich, Rebecca L; Miles, Adam R; Gale, Bruce K et al. (2009) Detergent screening of a G-protein-coupled receptor using serial and array biosensor technologies. Anal Biochem 386:98-104
Kieffer, Collin; Skalicky, Jack J; Morita, Eiji et al. (2008) Two distinct modes of ESCRT-III recognition are required for VPS4 functions in lysosomal protein targeting and HIV-1 budding. Dev Cell 15:62-73
Kim, Sunghwan; Pang, Hong-Bo; Kay, Michael S (2008) Peptide mimic of the HIV envelope gp120-gp41 interface. J Mol Biol 376:786-97

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