title: Transcriptional approaches to riboswitch studies.
abstract: Natural RNA sensors of small molecules (a.k.a. riboswitches) regulate numerous metabolic genes. In bacteria, these RNA elements control transcription termination and translation initiation by changing the folding pathway of nascent RNA upon direct binding of a metabolite. To identify and study riboswitches we used in vitro reconstituted solid-phase transcription elongation/termination system. This approach allows for direct monitoring of ligand binding and riboswitch functioning, establishing the working concentration of a ligand as a function of RNA polymerase speed, and also probing RNA structure of the riboswitch. Using this system we have been able to identify and characterize first several riboswitches including those involved in vitamin biosynthesis and sulfur metabolism. The system can be utilized to facilitate biochemical studies of riboswitches in general, i.e., to simplify analysis of riboswitches that are not necessarily involved in transcriptional control.
authors.affiliation: Department of Biochemistry, New York University Medical Center, New York, NY 10016, USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Mironov
authors.forename: Vitaly
authors.initials: V
authors.lastname: Epshtein
authors.forename: Evgeny
authors.initials: E
authors.lastname: Nudler
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D022661:RNA, Untranslated / Q000378:metabolism*; D038621:Regulatory Sequences, Ribonucleic Acid / Q000235:genetics; D013698:Templates, Genetic; D014158:Transcription, Genetic
chemical_list: D004269:DNA, Bacterial; D022661:RNA, Untranslated; D038621:Regulatory Sequences, Ribonucleic Acid; D012321:DNA-Directed RNA Polymerases