Human immunodeficiency virus type 1 (HIV-1) is a retrovirus that is the causative agent of human acquired immunodeficiency syndrome (AIDS). In retroviruses tRNA molecules are recruited as primers to initiate reverse transcription. In the case of HIV-1 a specific host cell tRNA, human tRNALys3, serves as the initiation primer. All three human tRNALys isoacceptors are selectively packaged into the HIV-1 virion. It has recently been shown that the tRNALys binding protein, human lysyl-tRNA synthetase (LysRS) is also packaged into HIV-1 virions and into viral-like particles formed from the HIV Gag protein alone. These data and recent pull-down assays support an interaction between human LysRS and Gag in vivo and in vitro. This proposal will explore this hypothesis further by quantitatively measuring and mapping the interaction in vitro. The goals will be accomplished by employing isothermal titration calorimetry (ITC), fluorescence polarization (FP), and protein footprinting techniques. Information gained from the proposed research will provide valuable insights into the molecular interaction between LysRS and Gag. Additionally, this information may be useful in the design of new therapeutic agents against AIDS.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32GM069339-01
Application #
6698508
Study Section
AIDS and Related Research 8 (AARR)
Program Officer
Marino, Pamela
Project Start
2003-08-11
Project End
2005-08-10
Budget Start
2003-08-11
Budget End
2004-08-10
Support Year
1
Fiscal Year
2003
Total Cost
$41,608
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Chemistry
Type
Other Domestic Higher Education
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
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Wei, Min; Yang, Yiliang; Niu, Meijuan et al. (2008) Inability of human immunodeficiency virus type 1 produced in murine cells to selectively incorporate primer formula. J Virol 82:12049-59
Kovaleski, Brandie J; Kennedy, Robert; Khorchid, Ahmad et al. (2007) Critical role of helix 4 of HIV-1 capsid C-terminal domain in interactions with human lysyl-tRNA synthetase. J Biol Chem 282:32274-9
Kovaleski, Brandie J; Kennedy, Robert; Hong, Minh K et al. (2006) In vitro characterization of the interaction between HIV-1 Gag and human lysyl-tRNA synthetase. J Biol Chem 281:19449-56