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Screening Assays to Identify Artificial glmS Ribozyme Activators

  • Christina E. Lünse
  • Günter Mayer
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1103)

Abstract

Ribsowitches are putative drug targets as they often regulate the expression of essential bacterial genes. This finding necessitates the development of suitable assays, at best high-throughput (HT) compatible, which allow the screening of compound libraries for riboswitch activation. Here, we describe a HT-compatible fluorescence-based screening assay employing a minimal core motif of the Bacillus subtilis glmS riboswitch and the metabolite-induced self-cleavage assay using the full-length glmS ribozyme of Staphylococcus aureus for the identification of artificial molecules activating this regulatory RNA.

Key words

glmS ribozyme Screening Ribozyme cleavage Riboswitch Fluorescence polarization 

References

  1. 1.
    Winkler WC, Nahvi A, Roth A, Collins JA, Breaker RR (2004) Control of gene expression by a natural metabolite-responsive ribozyme. Nature 428(6980):281–286PubMedCrossRefGoogle Scholar
  2. 2.
    Collins JA, Irnov I, Baker S, Winkler WC (2007) Mechanism of mRNA destabilization by the glmS ribozyme. Genes Dev 21(24):3356–3368PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Mayer G, Famulok M (2006) High-throughput-compatible assay for glmS riboswitch metabolite dependence. Chembiochem 7(4):602–604PubMedCrossRefGoogle Scholar
  4. 4.
    Lünse CE, Schmidt MS, Wittmann V, Mayer G (2011) Carba-sugars activate the glmS-riboswitch of Staphylococcus aureus. ACS Chem Biol 6(7):675–678PubMedCrossRefGoogle Scholar
  5. 5.
    Kladwang W, Hum J, Das R (2012) Ultraviolet shadowing of RNA can cause significant chemical damage in seconds. Sci Rep 2:517PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Christina E. Lünse
    • 1
  • Günter Mayer
    • 1
  1. 1.Life and Medical Sciences InstituteUniversity of BonnBonnGermany

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