Weigand, Julia E. ; Gottstein-Schmidtke, Sina R. ; Demolli, Shemsi ; Groher, Florian ; Duchardt-Ferner, Elke ; Wöhnert, Jens ; Suess, Beatrix (2014)
Sequence Elements Distal to the Ligand Binding Pocket Modulate the Efficiency of a Synthetic Riboswitch.
In: Chembiochem : a European journal of chemical biology, 15 (11)
Artikel, Bibliographie
Kurzbeschreibung (Abstract)
Synthetic riboswitches can serve as sophisticated genetic control devices in synthetic biology, regulating gene expression through direct RNA-ligand interactions. We analyzed a synthetic neomycin riboswitch, which folds into a stem loop structure with an internal loop important for ligand binding and regulation. It is closed by a terminal hexaloop containing a U-turn and a looped-out adenine. We investigated the relationship between sequence, structure, and biological activity in the terminal loop by saturating mutagenesis, ITC, and NMR. Mutants corresponding to the canonical U-turn fold retained biological activity. An improvement of stacking interactions in the U-turn led to an RNA element with slightly enhanced regulatory activity. For the first position of the U-turn motif and the looped out base, sequence-activity relationships that could not initially be explained on the basis of the structure of the aptamer-ligand complex were observed. However, NMR studies of these mutants revealed subtle relationships between structure and dynamics of the aptamer in its free or bound state and biological activity.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2014 |
| Autor(en): | Weigand, Julia E. ; Gottstein-Schmidtke, Sina R. ; Demolli, Shemsi ; Groher, Florian ; Duchardt-Ferner, Elke ; Wöhnert, Jens ; Suess, Beatrix |
| Art des Eintrags: | Bibliographie |
| Titel: | Sequence Elements Distal to the Ligand Binding Pocket Modulate the Efficiency of a Synthetic Riboswitch. |
| Sprache: | Englisch |
| Publikationsjahr: | 2014 |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Chembiochem : a European journal of chemical biology |
| Jahrgang/Volume einer Zeitschrift: | 15 |
| (Heft-)Nummer: | 11 |
| Kurzbeschreibung (Abstract): | Synthetic riboswitches can serve as sophisticated genetic control devices in synthetic biology, regulating gene expression through direct RNA-ligand interactions. We analyzed a synthetic neomycin riboswitch, which folds into a stem loop structure with an internal loop important for ligand binding and regulation. It is closed by a terminal hexaloop containing a U-turn and a looped-out adenine. We investigated the relationship between sequence, structure, and biological activity in the terminal loop by saturating mutagenesis, ITC, and NMR. Mutants corresponding to the canonical U-turn fold retained biological activity. An improvement of stacking interactions in the U-turn led to an RNA element with slightly enhanced regulatory activity. For the first position of the U-turn motif and the looped out base, sequence-activity relationships that could not initially be explained on the basis of the structure of the aptamer-ligand complex were observed. However, NMR studies of these mutants revealed subtle relationships between structure and dynamics of the aptamer in its free or bound state and biological activity. |
| Fachbereich(e)/-gebiet(e): | 10 Fachbereich Biologie 10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology") 10 Fachbereich Biologie > RNA Biochemie |
| Hinterlegungsdatum: | 01 Jul 2014 11:09 |
| Letzte Änderung: | 06 Nov 2020 07:13 |
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