The envelope glycoproteins, gp120 and gp41, of HIV are critical components of the infectious virus, that through specific interactions with the T4 molecule on the surface of CD4+ cells mediate virus attachment and entry. The N-terminus of gp41, which showns homology with paramyxovirus fusion-proteins and has been implicated in the process of virus/cell membrane fusion, is generated through proteolytic cleavage of the Pr160 env precursor during transport to the cell surface. Experiments in the avian retrovirus system show that in the absence of this cleavage, the glycoprotein is non-functional and the virus non-infectious. We propose therefore to employ site-directed mutagenesis, and site-specific antibodies to determine the specificity, site(s) and role of carboxypeptidase processing in cleavage/activation of the HIV env precursor. The Golgi-endopeptidase responsible for retroviral cleavage will be isolated and characterized with respect to proteinase class substrate/inhibitor specificity, and optimal reaction conditions. Monoclonal and polyclonal antibodies raised against the proteinase will be used for purification, subcellular localization and comparative studies. Limited peptide sequence on the proteinase will be obtained, such that synthetic oligonucleotide probes and antibodies can be used to probe cDNA libraries for gene cloning. Cloned gene sequences will be compared to other proteinase-super family members and used for structure-function studies on this important class of enzymes. The proposed research revolves around collaboration and interaction with other PEBRA investigators and thus will provide a major impetus for rapid dissemination of information and technologies.

Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of Alabama Birmingham
Department
Type
DUNS #
004514360
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Wilson, J E; Aulabaugh, A; Caligan, B et al. (1996) Human immunodeficiency virus type-1 reverse transcriptase. Contribution of Met-184 to binding of nucleoside 5'-triphosphate. J Biol Chem 271:13656-62
Dubay, J W; Dubay, S R; Shin, H J et al. (1995) Analysis of the cleavage site of the human immunodeficiency virus type 1 glycoprotein: requirement of precursor cleavage for glycoprotein incorporation. J Virol 69:4675-82
Spies, C P; Ritter Jr, G D; Mulligan, M J et al. (1994) Truncation of the cytoplasmic domain of the simian immunodeficiency virus envelope glycoprotein alters the conformation of the external domain. J Virol 68:585-91
Norris, J G; Tang, L P; Sparacio, S M et al. (1994) Signal transduction pathways mediating astrocyte IL-6 induction by IL-1 beta and tumor necrosis factor-alpha. J Immunol 152:841-50
Clark, S J; Kelen, G D; Henrard, D R et al. (1994) Unsuspected primary human immunodeficiency virus type 1 infection in seronegative emergency department patients. J Infect Dis 170:194-7
Benveniste, E N (1994) Cytokine circuits in brain. Implications for AIDS dementia complex. Res Publ Assoc Res Nerv Ment Dis 72:71-88
Byrne, J A; Stankovic, A K; Cooper, M D (1994) A novel subpopulation of primed T cells in the human fetus. J Immunol 152:3098-106
Bray, M; Prasad, S; Dubay, J W et al. (1994) A small element from the Mason-Pfizer monkey virus genome makes human immunodeficiency virus type 1 expression and replication Rev-independent. Proc Natl Acad Sci U S A 91:1256-60
Borrow, P; Lewicki, H; Hahn, B H et al. (1994) Virus-specific CD8+ cytotoxic T-lymphocyte activity associated with control of viremia in primary human immunodeficiency virus type 1 infection. J Virol 68:6103-10
Norris, J G; Benveniste, E N (1993) Interleukin-6 production by astrocytes: induction by the neurotransmitter norepinephrine. J Neuroimmunol 45:137-45

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