Abstract

The tatIII gene of human immunodeficiency virus (HIV), essential for viral replication, is involved in transactivation of HIV-LTR expression. We propose to introduce the HIV-LTR fused to the CAT gene into an adenovirus vector, and to use this system to analyze in detail the biochemical mechanisms involved in tatIII transactivation. Specifically, we intend to: 1. Use this recombinant HIV-LTR-adenovirus to infect HeLa cells expressing the tatIII gene (and control HeLa cells) and determine at what levels of regulation the tatIII protein acts on HIV-LTR gene expression. 2. Use this recombinant HIV-LTR-adenovirus to infect large numbers of cells in order to develop in vitro systems that mimic tatIII transactivation, allowing a dissection of the mechanisms at work. 3. Once the mode(s) of tatIII activation is known in this recombinant HIV-LTR-adenovirus system, to introduce the HIV- LTRCAT gene into pre-existing adenoviurses mutant in known functions (i.e., translational control, mRNA transport from the nucleus, transition from early to late phase of infection) in order to investigate the interplay of these well defined genes and the tatIII gene.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI025308-02
Application #
3138750
Study Section
(SRC)
Project Start
1987-09-30
Project End
1990-08-31
Budget Start
1988-09-01
Budget End
1989-08-31
Support Year
2
Fiscal Year
1988
Total Cost
Indirect Cost

  Institution

Name
Cold Spring Harbor Laboratory
Department
Type
DUNS #
065968786
City
Cold Spring Harbor
State
NY
Country
United States
Zip Code
11724

  Publications

Yang, X; Herrmann, C H; Rice, A P (1996) The human immunodeficiency virus Tat proteins specifically associate with TAK in vivo and require the carboxyl-terminal domain of RNA polymerase II for function. J Virol 70:4576-84
Herrmann, C H; Gold, M O; Rice, A P (1996) Viral transactivators specifically target distinct cellular protein kinases that phosphorylate the RNA polymerase II C-terminal domain. Nucleic Acids Res 24:501-8
Herrmann, C H; Rice, A P (1995) Lentivirus Tat proteins specifically associate with a cellular protein kinase, TAK, that hyperphosphorylates the carboxyl-terminal domain of the large subunit of RNA polymerase II: candidate for a Tat cofactor. J Virol 69:1612-20
Rhim, H; Rice, A P (1995) HIV-1 Tat protein is able to efficiently transactivate the HIV-2 LTR through a TAR RNA element lacking both dinucleotide bulge binding sites. Virology 206:673-8
Echetebu, C O; Rhim, H; Herrmann, C H et al. (1994) Construction and characterization of a potent HIV-2 Tat transdominant mutant protein. J Acquir Immune Defic Syndr 7:655-64
Rhim, H; Rice, A P (1994) Functional significance of the dinucleotide bulge in stem-loop1 and stem-loop2 of HIV-2 TAR RNA. Virology 202:202-11
Rhim, H; Rice, A P (1994) Exon2 of HIV-2 Tat contributes to transactivation of the HIV-2 LTR by increasing binding affinity to HIV-2 TAR RNA. Nucleic Acids Res 22:4405-13
Rhim, H; Echetebu, C O; Herrmann, C H et al. (1994) Wild-type and mutant HIV-1 and HIV-2 Tat proteins expressed in Escherichia coli as fusions with glutathione S-transferase. J Acquir Immune Defic Syndr 7:1116-21
Herrmann, C H; Rice, A P (1993) Specific interaction of the human immunodeficiency virus Tat proteins with a cellular protein kinase. Virology 197:601-8
Rice, A P; Wilson, E; Chan, F (1993) Limited proteolytic digestions identify common structural features of HIV-1 Tat proteins expressed during infection from alternatively spliced mRNAs. J Acquir Immune Defic Syndr 6:344-52

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