Genetic dissection of regulation by a repressing and novel activating corrinoid riboswitch enables engineering of synthetic riboswitches

In addition to proteins, microbes can use structured RNAs such as riboswitches for the important task of regulating gene expression. Riboswitches control gene expression by changing their structure in response to binding a small molecule and are widespread among bacteria. Here we determine the mecha...

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Bibliographic Details
Published in:mBio 2023-10, Vol.14 (5), p.e0158823
Main Authors: Procknow, Rebecca R, Kennedy, Kristopher J, Kluba, Maxwell, Rodriguez, Lesley J, Taga, Michiko E
Format: Article
Language:English
Subjects:
Bacillus subtilis
Bacteria - genetics
cobalamin
Coenzymes - metabolism
corrinoid
gene regulation
Genetic Engineering
Genetics and Molecular Biology
Research Article
riboswitch
Riboswitch - genetics
RNA biology
Vitamin B 12
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Summary:In addition to proteins, microbes can use structured RNAs such as riboswitches for the important task of regulating gene expression. Riboswitches control gene expression by changing their structure in response to binding a small molecule and are widespread among bacteria. Here we determine the mechanism of regulation in a riboswitch that responds to corrinoids-a family of coenzymes related to vitamin B . We report the alternative RNA secondary structures that couple corrinoid sensing with response in a repressing and novel activating corrinoid riboswitch. We then applied this knowledge to flipping the regulatory sign by constructing synthetic riboswitches that activate expression to a higher level than the natural one. In the process, we observed patterns in which sequence, in addition to structure, impacts function in paired RNA regions. The synthetic riboswitches we describe here have potential applications as biosensors.
ISSN:2150-7511
2150-7511
DOI:10.1128/mbio.01588-23