Abstract

The AIDS virus pol gene encodes three proteins that function as enzymes: a protease; the viral DNA polymerase (reverse transcriptase); and an endonuclease/integrase. Each of these proteins is a potential target for an inhibitor that could block virus replication. These proteins will be purified from bacteria that have been engineered to overproduce the AIDS virus pol gene products. These proteins will be used in biochemical analysis and for crystallization. The bacterial expression system will also be used for genetic analysis of the pol gene. Protein crystals formed in the presence and absence of specific ligands (including azido- thymidine triphosphate) will be used to determine the x-ray crystal structure using the isomorphous replacement method. The combined information obtained from genetic and biochemical studies and structure determination will provide a detailed understanding of each of these proteins and provide a basis for rational drug design.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI025321-03
Application #
3138779
Study Section
(SRC)
Project Start
1987-09-30
Project End
1990-08-31
Budget Start
1989-09-01
Budget End
1990-08-31
Support Year
3
Fiscal Year
1989
Total Cost
Indirect Cost

  Institution

Name
University of North Carolina Chapel Hill
Department
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599

  Publications

Resch, Wolfgang; Parkin, Neil; Watkins, Terri et al. (2005) Evolution of human immunodeficiency virus type 1 protease genotypes and phenotypes in vivo under selective pressure of the protease inhibitor ritonavir. J Virol 79:10638-49
Watkins, Terri; Resch, Wolfgang; Irlbeck, David et al. (2003) Selection of high-level resistance to human immunodeficiency virus type 1 protease inhibitors. Antimicrob Agents Chemother 47:759-69
Resch, Wolfgang; Ziermann, Rainer; Parkin, Neil et al. (2002) Nelfinavir-resistant, amprenavir-hypersusceptible strains of human immunodeficiency virus type 1 carrying an N88S mutation in protease have reduced infectivity, reduced replication capacity, and reduced fitness and process the Gag polyprotein precursor ab J Virol 76:8659-66
Pettit, Steve C; Henderson, Gavin J; Schiffer, Celia A et al. (2002) Replacement of the P1 amino acid of human immunodeficiency virus type 1 Gag processing sites can inhibit or enhance the rate of cleavage by the viral protease. J Virol 76:10226-33
Resch, W; Parkin, N; Stuelke, E L et al. (2001) A multiple-site-specific heteroduplex tracking assay as a tool for the study of viral population dynamics. Proc Natl Acad Sci U S A 98:176-81
Shehu-Xhilaga, M; Kraeusslich, H G; Pettit, S et al. (2001) Proteolytic processing of the p2/nucleocapsid cleavage site is critical for human immunodeficiency virus type 1 RNA dimer maturation. J Virol 75:9156-64
Wrobel, J A; Conrad, M J; Bloedon, E et al. (2000) Analysis of HIV type 1 reverse transcriptase: comparing sequences of viral isolates with mutational data. AIDS Res Hum Retroviruses 16:2049-54
Vaillancourt, M; Irlbeck, D; Smith, T et al. (1999) The HIV type 1 protease inhibitor saquinavir can select for multiple mutations that confer increasing resistance. AIDS Res Hum Retroviruses 15:355-63
Wrobel, J A; Chao, S F; Conrad, M J et al. (1998) A genetic approach for identifying critical residues in the fingers and palm subdomains of HIV-1 reverse transcriptase. Proc Natl Acad Sci U S A 95:638-45
Pettit, S C; Sheng, N; Tritch, R et al. (1998) The regulation of sequential processing of HIV-1 Gag by the viral protease. Adv Exp Med Biol 436:15-25

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