Rapid Depletion of Target Proteins Allows Identification of Coincident Physiological Responses
Abbreviated Journal Title
ESCHERICHIA-COLI STRAINS; MESSENGER-RNA DECAY; RIBOSOMAL-PROTEIN; POLYNUCLEOTIDE PHOSPHORYLASE; CLPXP DEGRADATION; STOP CODONS; S1; GENE; TRANSLATION; HELICASE; Microbiology
Targeted protein degradation is a powerful tool that can be used to create unique physiologies depleted of important factors. Current strategies involve modifying a gene of interest such that a degradation peptide is added to an expressed target protein and then conditionally activating proteolysis, either by expressing adapters, unmasking cryptic recognition determinants, or regulating protease affinities using small molecules. For each target, substantial optimization may be required to achieve a practical depletion, in that the target remains present at a normal level prior to induction and is then rapidly depleted to levels low enough to manifest a physiological response. Here, we describe a simplified targeted degradation system that rapidly depletes targets and that can be applied to a wide variety of proteins without optimizing target protease affinities. The depletion of the target is rapid enough that a primary physiological response manifests that is related to the function of the target. Using ribosomal protein Si as an example, we show that the rapid depletion of this essential translation factor invokes concomitant changes to the levels of several mRNAs, even before appreciable cell division has occurred.
Journal of Bacteriology
"Rapid Depletion of Target Proteins Allows Identification of Coincident Physiological Responses" (2012). Faculty Bibliography 2010s. 2354.