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Thermodynamic characterization of an engineered tetracycline-binding riboswitch.

Müller, Michael ; Weigand, Julia E. ; Weichenrieder, Oliver ; Suess, Beatrix (2006)
Thermodynamic characterization of an engineered tetracycline-binding riboswitch.
In: Nucleic acids research, 34 (9)
Artikel, Bibliographie

Kurzbeschreibung (Abstract)

Riboswitches reflect a novel concept in gene regulation that is particularly suited for technological adaptation. Therefore, we characterized thermodynamically the ligand binding properties of a synthetic, tetracycline (tc)-binding RNA aptamer, which regulates gene expression in a dose-dependent manner when inserted into the untranslated region of an mRNA. In vitro, one molecule of tc is bound by one molecule of partially pre-structured and conformationally homogeneous apo-RNA. The dissociation constant of 770 pM, as determined by fluorimetry, is the lowest reported so far for a small molecule-binding RNA aptamer. Additional calorimetric analysis of RNA point mutants and tc derivatives identifies functional groups crucial for the interaction and including their respective enthalpic and entropic contributions we can propose detailed structural and functional roles for certain groups. The conclusions are consistent with mutational analyses in vivo and support the hypothesis that tc-binding reinforces the structure of the RNA aptamer, preventing the scanning ribosome from melting it efficiently.

Typ des Eintrags: Artikel
Erschienen: 2006
Autor(en): Müller, Michael ; Weigand, Julia E. ; Weichenrieder, Oliver ; Suess, Beatrix
Art des Eintrags: Bibliographie
Titel: Thermodynamic characterization of an engineered tetracycline-binding riboswitch.
Sprache: Englisch
Publikationsjahr: 2006
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Nucleic acids research
Jahrgang/Volume einer Zeitschrift: 34
(Heft-)Nummer: 9
Kurzbeschreibung (Abstract):

Riboswitches reflect a novel concept in gene regulation that is particularly suited for technological adaptation. Therefore, we characterized thermodynamically the ligand binding properties of a synthetic, tetracycline (tc)-binding RNA aptamer, which regulates gene expression in a dose-dependent manner when inserted into the untranslated region of an mRNA. In vitro, one molecule of tc is bound by one molecule of partially pre-structured and conformationally homogeneous apo-RNA. The dissociation constant of 770 pM, as determined by fluorimetry, is the lowest reported so far for a small molecule-binding RNA aptamer. Additional calorimetric analysis of RNA point mutants and tc derivatives identifies functional groups crucial for the interaction and including their respective enthalpic and entropic contributions we can propose detailed structural and functional roles for certain groups. The conclusions are consistent with mutational analyses in vivo and support the hypothesis that tc-binding reinforces the structure of the RNA aptamer, preventing the scanning ribosome from melting it efficiently.

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: 22 Feb 2012 10:35
Letzte Änderung: 06 Nov 2020 07:24
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