Abstract

The structural information obtained through crystallographic studies on RSV and HIV-1 proteases will guide the present study in attempts to examine the role of critical residues that determine the substrate selection and catalytic efficiency. Attempts to change the specificity of one PR into the other for both heterologous and homologous substrates will be undertaken by site-directed mutagenesis and tested by biochemical characterization of purified enzymes. The effect of PR subunit symmetry for optimal catalytic efficiencies will be examined by selective mutagenesis of individual subunits of the PR homodimer. The information gathered from these experiments would be used to construct a """"""""designer protease"""""""" to act on other than normal target sites on HIV-1 RT. Finally, altered proteases will be tested in vivo , for further understanding of their role in biological processes involved in virus assembly and reproduction.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA052047-07
Application #
2700440
Study Section
AIDS and Related Research Study Section 3 (ARRC)
Program Officer
Cole, John S
Project Start
1992-07-01
Project End
1999-02-28
Budget Start
1998-05-01
Budget End
1999-02-28
Support Year
7
Fiscal Year
1998
Total Cost
Indirect Cost

  Institution

Name
Case Western Reserve University
Department
Biochemistry
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106

  Publications

Chen, Aiping; Weber, Irene T; Harrison, Robert W et al. (2006) Identification of amino acids in HIV-1 and avian sarcoma virus integrase subsites required for specific recognition of the long terminal repeat Ends. J Biol Chem 281:4173-82
Medina, Gisselle; Zhang, Yongjun; Tang, Yi et al. (2005) The functionally exchangeable L domains in RSV and HIV-1 Gag direct particle release through pathways linked by Tsg101. Traffic 6:880-94
Vana, Marcy L; Chen, Aiping; Boross, Peter et al. (2005) Mutations affecting cleavage at the p10-capsid protease cleavage site block Rous sarcoma virus replication. Retrovirology 2:58
Vana, Marcy L; Tang, Yi; Chen, Aiping et al. (2004) Role of Nedd4 and ubiquitination of Rous sarcoma virus Gag in budding of virus-like particles from cells. J Virol 78:13943-53
Johnson, Michael; Morris, Shannon; Chen, Aiping et al. (2004) Selection of functional mutations in the U5-IR stem and loop regions of the Rous sarcoma virus genome. BMC Biol 2:8
Brin, Elena; Leis, Jonathan (2002) HIV-1 integrase interaction with U3 and U5 terminal sequences in vitro defined using substrates with random sequences. J Biol Chem 277:18357-64
Brin, Elena; Leis, Jonathan (2002) Changes in the mechanism of DNA integration in vitro induced by base substitutions in the HIV-1 U5 and U3 terminal sequences. J Biol Chem 277:10938-48
Morris, Shannon; Johnson, Michael; Stavnezer, Ed et al. (2002) Replication of avian sarcoma virus in vivo requires an interaction between the viral RNA and the TpsiC loop of the tRNA(Trp) primer. J Virol 76:7571-7
Mayo, Keith; Vana, Marcy L; McDermott, Jason et al. (2002) Analysis of Rous sarcoma virus capsid protein variants assembled on lipid monolayers. J Mol Biol 316:667-78
VerPlank, L; Bouamr, F; LaGrassa, T J et al. (2001) Tsg101, a homologue of ubiquitin-conjugating (E2) enzymes, binds the L domain in HIV type 1 Pr55(Gag). Proc Natl Acad Sci U S A 98:7724-9

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