The Rev transactivator of human immunodeficiency virus type 1 (HIV-1) is a posttranscriptional regulatory protein that is essential for replication of the virus. Localized within the nuclei of infected cells, Rev acts through a poorly understood mechanism to allow cytoplasmic accumulation of certain incompletely spliced HIV-1 mRNAs that would otherwise remain sequestered in the nucleus. Because these mRNAs code for the major viral structural proteins, HIV- 1 proviruses that lack a functional rev gene are unable to produce new infectious virions. Rev is therefore of interest not only because of its unprecedented regulatory effect, but also as a promising target for antiretroviral therapy. Using techniques of molecular genetics, we are investigating the functional architecture and mechanism of action of Rev, searching for potential means of inhibiting its activity. Mutations are introduced systematically into Rev and its cis-acting target element in the viral genome, and the effects of these mutations are then characterized using a transient transfection assay. Our studies have identified, and will now further characterize, three critical peptide domains that mediate interactions of Rev monomers with each other, with putative cellular cofactors, and with the target RNAs. Essential features of each region will be mapped in detail to provide a basis for rational design of inhibitors, and we will search for equivalent domains in Rev-like proteins from other retroviruses and in selected cellular proteins. We will also pursue the observation that modified forms of these regions can block activity of wild-type Rev, as a possible approach to gene therapy. Using Rev fusion proteins that have altered target specificity, we will dissect the minimal requirements for this pathway of viral transactivation in vivo. We will also investigate how changes in the physiologic state of an infected T lymphocyte can modulate Rev activity. The information to be gained in these studies will be invaluable in designing novel pharmacologic or genetic strategies for inhibiting Rev, which could be used to maintain HIV-1 latency and slow the progression of disease in infected individuals.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI029313-04
Application #
3144054
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Project Start
1990-03-01
Project End
1997-02-28
Budget Start
1993-03-01
Budget End
1994-02-28
Support Year
4
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of California San Francisco
Department
Type
Schools of Medicine
DUNS #
073133571
City
San Francisco
State
CA
Country
United States
Zip Code
94143
Clever, J L; Eckstein, D A; Parslow, T G (1999) Genetic dissociation of the encapsidation and reverse transcription functions in the 5' R region of human immunodeficiency virus type 1. J Virol 73:101-9
Clever, J L; Parslow, T G (1997) Mutant human immunodeficiency virus type 1 genomes with defects in RNA dimerization or encapsidation. J Virol 71:3407-14
Lochrie, M A; Waugh, S; Pratt Jr, D G et al. (1997) In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein. Nucleic Acids Res 25:2902-10
Clever, J L; Wong, M L; Parslow, T G (1996) Requirements for kissing-loop-mediated dimerization of human immunodeficiency virus RNA. J Virol 70:5902-8
Mujeeb, A; Parslow, T G; Yuan, Y C et al. (1996) Aqueous solution structure of a hybrid lentiviral Tat peptide and a model of its interaction with HIV-1 TAR RNA. J Biomol Struct Dyn 13:649-60
Hunter, J J; Bond, B L; Parslow, T G (1996) Functional dissection of the human Bc12 protein: sequence requirements for inhibition of apoptosis. Mol Cell Biol 16:877-83
Lee, L C; Hunter, J J; Mujeeb, A et al. (1996) Evidence for alpha-helical conformation of an essential N-terminal region in the human Bcl2 protein. J Biol Chem 271:23284-8
Hunter, J J; Parslow, T G (1996) A peptide sequence from Bax that converts Bcl-2 into an activator of apoptosis. J Biol Chem 271:8521-4
Clever, J; Sassetti, C; Parslow, T G (1995) RNA secondary structure and binding sites for gag gene products in the 5' packaging signal of human immunodeficiency virus type 1. J Virol 69:2101-9
Mujeeb, A; Bishop, K; Peterlin, B M et al. (1994) NMR structure of a biologically active peptide containing the RNA-binding domain of human immunodeficiency virus type 1 Tat. Proc Natl Acad Sci U S A 91:8248-52

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