Site-specific photolabile roadblocks for the study of transcription elongation in biologically complex systems.

ABSTRACT: Transcriptional pausing is crucial for the timely expression of genetic information. Biochemical methods quantify the half-life of paused RNA polymerase (RNAP) by monitoring restarting complexes across time. However, this approach may produce apparent half-lives that are longer than true pause escape rates in biological contexts where multiple consecutive pause sites are present. We show here that the 6-nitropiperonyloxymethyl (NPOM) photolabile group provides an approach to monitor transcriptional pausing in biological systems containing multiple pause sites. We validate our approach using the well-studied his pause and show that an upstream RNA sequence modulates the pause half-life. NPOM was also used to study a transcriptional region within the Escherichia coli thiC riboswitch containi

SUBMITTER: Nadon JF 

PROVIDER: S-EPMC9098449 | biostudies-literature | 2022 May

REPOSITORIES: biostudies-literature