The broad objective of this proposal is to elucidate the mechanism of HIV-1 reverse transcription at the molecular level. The reverse transcription of the HIV-1 viral genome is a critical step in the replication cycle of the virus which is only poorly understood. At present, there is no system available for the thorough study of the biochemical processes required for HIV-1 reverse transcription. In order to approach this problem, we plan to express and characterize HIV-1 proteins required for reverse transcription. Specifically, enzymatically active RT from E. coli will be characterized, and techniques for the complete biochemical purification will be developed. Additional virion proteins (eg. gag gene products, endonuclease, accessory gene products) which might also be required in reverse transcription will be expressed using a similar approach. We will develop a reconstituted, in vitro system which accurately reflects HIV-1 reverse transcription. The expressed HIV-1 RT, and template RNA, generated from in vitro transcribed infectious HIV-1 proviral DNA, will be used to develop an in vitro system in which we can define the molecular events of HIV-1 reverse transcription. The specific structural features of HIV-1 RT will be correlated with function in reverse transcription of the HIV-1 genome. DNA mutagenesis will be used to create mutant RTs to elucidate structural features of this protein that are required for activity in the in vitro reverse transcription system. Finally, mutant RT will be tested for the capacity to interfere with reverse transcription in vitro and inhibit HIV-1 replication in susceptible tissue culture cells. An important component in the development of a rational approach for the control of AIDS is to elucidate the events of viral replication. The long term goal of this project, to understand the molecular details of the mechanism of HIV-1 reverse transcription, will provide a foundation for the development of novel methods in which to more effectively control virus replication.

Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
1989
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
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
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
Piatak Jr, M; Saag, M S; Yang, L C et al. (1993) Determination of plasma viral load in HIV-1 infection by quantitative competitive polymerase chain reaction. AIDS 7 Suppl 2:S65-71

Showing the most recent 10 out of 58 publications