TY  - JOUR
A1  - Müller, Michael
A1  - Weigand, Julia E.
A1  - Weichenrieder, Oliver
A1  - Süß, Beatrix
T1  - Thermodynamic characterization of an engineered tetracycline-binding riboswitch
T2  - Nucleic acids research
N2  - 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.
Y1  - 2006
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/26383
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-263830
UR  - http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1463898/
SN  - 1362-4962
SN  - 0305-1048
N1  - (c) The Author 2006. Published by Oxford University Press. All rights reserved. The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org
VL  - 34
IS  - 9
SP  - 2607
EP  - 2617
PB  - Oxford Univ. Press
CY  - Oxford
ER  -