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- Title
Protein-dependent ribozymes report molecular interactions in real time.
- Authors
Hartig, Jörg S.; Najafi-Shoushtari, S. Hani; Grüne, Imke; Yan, Amy; Ellington, Andrew D.; Famulok, Michael
- Abstract
Most approaches to monitoring interactions between biological macromolecules require large amounts of material, rely upon the covalent modification of an interaction partner, or are not amenable to real-time detection. We have developed a generalizable assay system based on interactions between proteins and reporter ribozymes. The assay can be configured in a modular fashion to monitor the presence and concentration of a protein or of molecules that modulate protein function. We report two applications of the assay: screening for a small molecule that disrupts protein binding to its nucleic acid target and screening for protein-protein interactions. We screened a structurally diverse library of antibiotics for small molecules that modulate the activity of HIV-1 Revresponsive ribozymes by binding to Rev. We identified an inhibitor that subsequently inhibited HIV-1 replication in cells. A simple format switch allowed reliable monitoring of domain-specific interactions between the bloodclotting factor thrombin and its protein partners. The rapid identification of interactions between proteins or of compounds that disrupt such interactions should have substantial utility for the drug-discovery process.
- Subjects
CATALYTIC RNA; BIOMACROMOLECULES
- Publication
Nature Biotechnology, 2002, Vol 20, Issue 7, p717
- ISSN
1087-0156
- Publication type
Article
- DOI
10.1038/nbt0702-717