Exploiting preQ1 riboswitches to regulate ribosomal frameshifting

Chien-hung Yu, Jinghui Luo, Dirk Iwata-Reuyl, René C.L. Olsthoorn

Research output: Contribution to journalArticlepeer-review

10 Citations (Scopus)

Abstract

Knowing the molecular details of the interaction between riboswitch aptamers and their corresponding metabolites is important to understand gene expression. Here we report on a novel in vitro assay to study preQ1 riboswitch aptamers upon binding of 7-Aminomethyl-7-deazaguanine (preQ 1). The assay is based on the ability of the preQ1 aptamer to fold, upon ligand binding, into a pseudoknotted structure that is capable of stimulating -1 ribosomal frameshifting (-1 FS). Aptamers from three different species were found to induce between 7% and 20% of -1 FS in response to increasing preQ1 levels, whereas preQ1 analogues were 100-1000-fold less efficient. In depth mutational analysis of the Fusobacterium nucleatum aptamer recapitulates most of the structural details previously identified for preQ1 aptamers from other bacteria by crystallography and/or NMR spectroscopy. In addition to providing insight into the role of individual nucleotides of the preQ1 riboswitch aptamer in ligand binding, the presented system provides a valuable tool to screen small molecules against bacterial riboswitches in a eukaryotic background.

Original languageEnglish
Pages (from-to)733-740
Number of pages8
JournalACS Chemical Biology
Volume8
Issue number4
DOIs
Publication statusPublished - 2013 Apr 19

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine

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