Protein-dependent ribozymes report molecular interactions in real time

Jörg S. Hartig, Hani Najafi, Imke Grüne, Amy Yan, Andrew D. Ellington, Michael Famulok

Research output: Contribution to journalArticle

129 Citations (Scopus)

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 blood-clotting 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.

Original languageEnglish
Pages (from-to)717-722
Number of pages6
JournalNature Biotechnology
Volume20
Issue number7
DOIs
Publication statusPublished - 2002
Externally publishedYes

Fingerprint

Catalytic RNA
Molecular interactions
Proteins
Assays
Molecules
HIV-1
Screening
Small Molecule Libraries
Blood Coagulation Factors
Monitoring
Nucleic acids
Blood Coagulation
Antibiotics
Drug Discovery
Macromolecules
Protein Binding
Thrombin
Nucleic Acids
Blood
Switches

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Biomedical Engineering
  • Molecular Medicine

Cite this

Hartig, J. S., Najafi, H., Grüne, I., Yan, A., Ellington, A. D., & Famulok, M. (2002). Protein-dependent ribozymes report molecular interactions in real time. Nature Biotechnology, 20(7), 717-722. https://doi.org/10.1038/nbt0702-717

Protein-dependent ribozymes report molecular interactions in real time. / Hartig, Jörg S.; Najafi, Hani; Grüne, Imke; Yan, Amy; Ellington, Andrew D.; Famulok, Michael.

In: Nature Biotechnology, Vol. 20, No. 7, 2002, p. 717-722.

Research output: Contribution to journalArticle

Hartig, JS, Najafi, H, Grüne, I, Yan, A, Ellington, AD & Famulok, M 2002, 'Protein-dependent ribozymes report molecular interactions in real time', Nature Biotechnology, vol. 20, no. 7, pp. 717-722. https://doi.org/10.1038/nbt0702-717
Hartig, Jörg S. ; Najafi, Hani ; Grüne, Imke ; Yan, Amy ; Ellington, Andrew D. ; Famulok, Michael. / Protein-dependent ribozymes report molecular interactions in real time. In: Nature Biotechnology. 2002 ; Vol. 20, No. 7. pp. 717-722.
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