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Nanomanipulation of Single RNA Molecules by Optical Tweezers
Published on: August 20, 2014
Mutational analysis of structural elements in a class-I cyclic di-GMP riboswitch to elucidate its regulatory
Saki Inuzuka1, Kei-Ichiro Nishimura2, Hitoshi Kakizawa1
1Department of Chemistry, Graduate School of Science and Engineering, University of Toyama, Toyama 930-8555, Japan.
Abstract:
The Vc2 riboswitch possesses an aptamer domain belonging to the class-I c-di-GMP riboswitch family. This domain has been analysed and the molecular mechanism by which it recognizes the c-di-GMP ligand has been elucidated. On the other hand, the regulatory mechanism of the full-length Vc2 riboswitch to control its downstream open reading frame (ORF) remains largely unknown. In this study, we performed in vivo reporter assays and in vitro biochemical analyses of the full-length riboswitch and its aptamer domain. We evaluated the results of in vivo and in vitro analyses to elucidate the regulatory mechanism of the Vc2 riboswitch. The present results suggest that recognition of c-di-GMP ligand by the Vc2 riboswitch aptamer domain downregulates expression of its downstream ORF primarily at the translational level.
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