This project aims at creating and using the tools to screen large libraries of chemical compounds to find and evaluate drug candidates for three targets in HIV-1. These are (1) the RNAnucleocapsid (NC) protein complex, (2) the TAR RNA-Tat protein-CyclinT1 protein complex, and (3) the Vif protein. No inhibitors of these targets are available in current AIDS therapies, in phase I of this project we succeeded in creating a molecular switch that changes its state in the presence of NC, and switches back in the presence of an inhibitor that mimics the viral RNA packaging signal. The inhibitor causes an increase in fluorescence within seconds that is easily monitored in high throughput screens (HTS) of candidate drugs. It is easy and unambiguous to verify that the complex occurs with a 1:1 ratio of protein and switch components (or not), and to determine the affinity constant, K/d. We propose limited prototyping and to create HTS- and Kdkits that others can use to discover and refine drug candidates to interrupt NC, Tat, and Vif function in HIV-I. In the future, similar assays will target all fifteen HIV proteins with new drugs.
| DeCiantis, Christopher L; Jensen, Danielle K; Hudson, Bruce S et al. (2007) A nucleic acid switch triggered by the HIV-1 nucleocapsid protein. Biochemistry 46:9164-73 |
