Screening assays to identify artificial glmS ribozyme activators

Methods Mol Biol. 2014;1103:199-209. doi: 10.1007/978-1-62703-730-3_15.

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.

MeSH terms

  • Bacillus subtilis / genetics
  • Bacterial Proteins / genetics*
  • Gene Expression Regulation, Bacterial
  • High-Throughput Screening Assays*
  • Molecular Biology / methods
  • Nucleic Acid Conformation
  • RNA, Catalytic / genetics*
  • Riboswitch / genetics
  • Staphylococcus aureus / genetics

Substances

  • Bacterial Proteins
  • RNA, Catalytic
  • Riboswitch
  • component S, glutamate mutase protein, Bacteria