The purpose of this research project is to study thoroughly the molecular and biological properties of two proteins that play key roles in the life cycle of human immunodeficiency virus (HIV-the AIDS virus). The first is the enzyme RNA-dependent DNA polymerase (reverse transcriptase-RT), and the second is the integration protein. These proteins are involved in the process of transcribing the viral RNA into DNA and in the subsequent integration of the viral DNA into the hose genome. The gene segments encoding these proteins were cloned by us in E. coli expression vectors. Bacterial strains that contain these expression plasmids produce substantial amounts of the two proteins. Both proteins closely approximate the corresponding viral proteins in the primary amino acid sequence. Purification from the bacteria of large amounts of these HIV proteins (which, at least in the case of RT, have been shown to be fully enzymatically active) will allow detailed biochemical and structural studies and provide a clearer understanding of the functions of the proteins. The information obtained should be useful in screening and designing drugs that inhibit the activities of the proteins, and should be helpful in the development of new drugs effective against AIDS. The E. coli system is suitable for the generation of mutations of the HIV proteins and the subsequent analyses of the effects of these mutations on the catalytic activities and drug sensitivity of the proteins. We propose to generate and screen for mutations of RT that might evade the present drugs used against AIDS, which could be helpful in developing drugs effective against mutated forms of HIV RT that might arise in the future.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI027035-03
Application #
3141087
Study Section
Special Emphasis Panel (ARR (V1))
Project Start
1989-09-30
Project End
1992-07-31
Budget Start
1991-08-01
Budget End
1992-07-31
Support Year
3
Fiscal Year
1991
Total Cost
Indirect Cost
Name
Tel Aviv University
Department
Type
DUNS #
City
Tel Aviv
State
Country
Israel
Zip Code
69978
Rubinek, T; Loya, S; Shaharabany, M et al. (1994) The catalytic properties of the reverse transcriptase of the lentivirus equine infectious anemia virus. Eur J Biochem 219:977-83
Loya, S; Bakhanashvili, M; Tal, R et al. (1994) Enzymatic properties of two mutants of reverse transcriptase of human immunodeficiency virus type 1 (tyrosine 181-->isoleucine and tyrosine 188-->leucine), resistant to nonnucleoside inhibitors. AIDS Res Hum Retroviruses 10:939-46
Hizi, A; Tal, R; Shaharabany, M et al. (1993) Specific inhibition of the reverse transcriptase of human immunodeficiency virus type 1 and the chimeric enzymes of human immunodeficiency virus type 1 and type 2 by nonnucleoside inhibitors. Antimicrob Agents Chemother 37:1037-42
Shaharabany, M; Rice, N R; Hizi, A (1993) Expression and mutational analysis of the reverse transcriptase of the lentivirus equine infectious anemia virus. Biochem Biophys Res Commun 196:914-20
Bakhanashvili, M; Hizi, A (1993) The fidelity of the reverse transcriptases of human immunodeficiency viruses and murine leukemia virus, exhibited by the mispair extension frequencies, is sequence dependent and enzyme related. FEBS Lett 319:201-5
Bakhanashvili, M; Hizi, A (1993) Fidelity of DNA synthesis exhibited in vitro by the reverse transcriptase of the lentivirus equine infectious anemia virus. Biochemistry 32:7559-67
Hizi, A; Shaharabany, M; Tal, R et al. (1992) The effects of cysteine mutations on the reverse transcriptases of human immunodeficiency virus types 1 and 2. J Biol Chem 267:1293-7
Hizi, A; Shaharabany, M (1992) Functional analysis of novel selective mutants of the reverse transcriptase of human immunodeficiency virus type 1. J Biol Chem 267:18255-8
Bakhanashvili, M; Hizi, A (1992) Fidelity of the reverse transcriptase of human immunodeficiency virus type 2. FEBS Lett 306:151-6
Loya, S; Tal, R; Hughes, S H et al. (1992) The effects of cysteine mutations on the catalytic activities of the reverse transcriptase of human immunodeficiency virus type-1. J Biol Chem 267:13879-83

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