Single-molecule microscopy image data and analysis files for “A translational riboswitch coordinates nascent transcription–translation coupling.”

These data were generated to study the dynamics of RNAP-ribosome interactions on a nascent mRNA with a preQ1-sensing translational riboswitch in its 5’ untranslated region (UTR). Using single-molecule fluorescence co-localization, we monitored direct transient binding of 30S ribosomal subunit to the individual nascent mRNA molecules in surface-immobilized paused elongation complexes (PECs). Also, using a novel protein-induced fluorescence enhancement assay we monitored the real-time transcription rate of RNA polymerase (RNAP) under different experimental conditions. The DNA template including the preQ1 riboswitch from B. anthracis under the control of the T7A1 promoter was cloned into pUC19 plasmid. Transcription templates for in vitro transcription were generated by PCR.