Efficient quantitative monitoring of translational initiation by RelE cleavage.

ABSTRACT: The sequences of the 5' untranslated regions (5'-UTRs) of mRNA alter gene expression across domains of life. Transcriptional modulators can be easily assayed through transcription termination, but translational regulators often require indirect, laborious methods. We have leveraged RelE's ribosome-dependent endonuclease activity to develop a quantitative assay to monitor translation initiation of cis-regulatory mRNAs. RelE cleavage accurately reports ligand-dependent changes in ribosome association for two translational riboswitches and provides quantitative information about each switch's sensitivity and range of response. RelE accurately reads out sequence-driven changes in riboswitch specificity and function and is quantitatively dependent upon ligand concentration. RelE cleavage simila

SUBMITTER: Focht CM 

PROVIDER: S-EPMC9561414 | biostudies-literature | 2022 Oct

REPOSITORIES: biostudies-literature