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Single-molecule FRET studies on the cotranscriptional folding of a thiamine pyrophosphate riboswitch.


ABSTRACT: Because RNAs fold as they are being synthesized, their transcription rate can affect their folding. Here, we report the results of single-molecule fluorescence studies that characterize the ligand-dependent cotranscriptional folding of the Escherichia coli thiM riboswitch that regulates translation. We found that the riboswitch aptamer folds into the "off" conformation independent of its ligand, but switches to the "on" conformation during transcriptional pausing near the translational start codon. Ligand binding maintains the riboswitch in the off conformation during transcriptional pauses. We expect our assay will permit the controlled study of the two main physical mechanisms that regulate cotranscriptional folding: transcriptional pausing and transcriptional speed.

SUBMITTER: Uhm H 

PROVIDER: S-EPMC5777041 | biostudies-literature | 2018 Jan

REPOSITORIES: biostudies-literature

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Single-molecule FRET studies on the cotranscriptional folding of a thiamine pyrophosphate riboswitch.

Uhm Heesoo H   Kang Wooyoung W   Ha Kook Sun KS   Kang Changwon C   Hohng Sungchul S  

Proceedings of the National Academy of Sciences of the United States of America 20171226 2


Because RNAs fold as they are being synthesized, their transcription rate can affect their folding. Here, we report the results of single-molecule fluorescence studies that characterize the ligand-dependent cotranscriptional folding of the <i>Escherichia coli thiM</i> riboswitch that regulates translation. We found that the riboswitch aptamer folds into the "off" conformation independent of its ligand, but switches to the "on" conformation during transcriptional pausing near the translational st  ...[more]

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