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Roles of Mg2+ in TPP-dependent riboswitch
Authors:Yamauchi Takahiro  Miyoshi Daisuke  Kubodera Takafumi  Nishimura Akira  Nakai Susumu  Sugimoto Naoki
Institution:Frontier Institute for Biomolecular Engineering, Research (FIBER), Konan University, Higashinada-ku, Kobe, Japan.
Abstract:We quantified the effect of Mg(2+) on thiamine pyrophosphate (TPP) binding to TPP-dependent thiA riboswitch RNA. The association constant of TPP binding to the riboswitch at 20 degrees C increased from 1.2 x 10(6) to 50 x 10(6) M(-1) as the Mg(2+) concentration increased from 0 to 1 mM. Furthermore, circular dichroic spectra under various conditions showed that 1 mM Mg(2+) induced a local structural change of the riboswitch, which might be pivotal for TPP binding. These results indicate that a physiological concentration of Mg(2+) can regulate TPP binding to the thiA riboswitch.
Keywords:TPP  thiamine pyrophosphate  Ka  association constant  5′-UTR  5′-untranslated region  PCR  polymerase chain reaction  PAGE  polyacrylamide gel electrophoresis  CD  circular dichroism  UV  ultra violet
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