Cellular transcription of DNA-encoded information into RNA is a complex, highly regulated process carried out by multi-subunit RNA polymerases (RNAPs). Transcription by RNAPs is not a continuous process, but is instead interrupted by DNA sequence-dependent pauses, during which the polymerase remains bound to the nucleic acids without active RNA synthesis occurring. Transcriptional pausing plays crucial regulatory roles in both prokaryotes and eukaryotes. Pausing in bacteria, for instance, allows synchronization of transcription and translation, creates time windows for transcription regulators to bind the elongating transcription complex, serves as an initial step in transcription termination, and promotes the proper folding of nascent RNA transcripts. Despite its importance in regulation of gene expression, the pausing phenomenon is not completely understood at the molecular level. Here, I will describe insights into the underlying mechanisms of RNAP pausing gained from biochemical and structural studies, including preliminary efforts at resolving conformational intermediates of RNA polymerase <i>en route</i> to a paused state using time-resolved single-particle cryoEM.

Pausing by bacterial RNA polymerase: Mechanistic insights from structural and biochemical studies

2019 Fall National Meeting

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