UC San Diego

Protein production is tightly regulated by post transcription factors to control internal cellular elements and adapt themselves to the surrounding environment, such as the nutrient availability, temperature and oxygen absence. Translational elongation is one of the regulators for protein production. The mRNA localization to mitochondria during translational elongation has been reported to induce the increase in its protein production. The metabolic shift is a key factor for ATP synthase genes to localize its mRNAs to mitochondria during respiration to escalate ATP productions under oxidative phosphorylation. In the respiration, some ATP synthase genes are reported to induce the localization switching metabolism from fermentation to respiration. We have been investigating if slowing translational elongation can improve the protein production in conditionally localized mRNAs. Here we found that the ATP2 and ATP4 in yeast cells have decent increases in mRNA levels upon translational elongation slowing. In addition to it, the promoter has a certain effect in protein induction under respiration, but promoter combined with ORF has the stronger effect in it. To identify genes involved in mRNA localization-dependent gene expression, we have constructed the design of CRISPRi screening with a group of gRNAs targets various promoters in yeast cells in combination with a MCP- MS2 system to target a reporter mRNA to the mitochondria.