Regulation of gene expression determines the functional yields of recombinant proteins in the common eukaryotic host <i>Saccharomyces cerevisiae</i>. Despite numerous advances in yeast genetics and molecular biology, current methods to control recombinant protein expression in yeast cultures are limited to induction of a few select promoters by small molecules. Here, we present a novel approach to activate transcription in yeast by reconstitution of peptide signals using split intein-mediated reactions. These reactions are designed to use inactive fusion proteins to produce functional α-factor peptide analogues that bind onto Ste2p, a native yeast G protein-coupled receptor, and activate yeast mating response and transcriptional induction. By integrating this native mating signaling pathway with heterologous gene expression cassettes, we can control timing and level of induction by applying our split intein fusion proteins. This synthetic control strategy will enable regulation of yeast functions in a wide range of applications, including recombinant protein production and consolidated bioprocessing.

Transcriptional activation of yeast by split intein-mediated reconstitution of synthetic peptide signals

249th National Meeting (2015)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.