Stress responses to heterologous membrane protein expression in Escherichia coli
The stress response of E. coli to the expression of two recombinant membrane proteins, the E. coli AAA+ protease FtsH and the human G-protein coupled receptor CB1, was examined using several members of a promoter-GFP library. Several genes from the heat-shock and envelope stress regulons (rpoH, clpP, lon, and ftsH) were strongly induced by expression of either membrane protein. Flow cytometry was used to monitor the real-time dynamics of the transcription of these reporter genes in response to membrane protein expression. Co-expression of CB1 and FtsH led to an additive response in these four reporter genes suggesting that the stresses may be occurring via different physiological mechanisms.
KeywordsG protein-coupled receptors Heterologous protein expression Sigma factor Stress response
This research was supported by Princeton University startup funds and by the Lidow Senior Thesis fund. The authors thank Dr. Eva-Maria Strauch (University of Washington) for comments on the manuscript.
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