Expression in E. coli of hyper- and hypo-amyloidogenic RepA-WH1 variants

RepA-WH1 is a synthetic bacterial prionoid, i.e., a protein that aggregates as amyloid in bacteria leading to cell death The purpose of this approach was to outline the pathways of cytotoxicity linked to RepA-WH1 expression from the set of genes induced/repressed in an Escherichia coli strain with reduced genome (MDS42) Transcription profiles in cells expressing the mildly-citotoxic RepA-WH1(∆N37)-mCherry aggregates were subtracted from the profiles for cells expressing the acutely-cytotoxic A31V amyloid aggregates. This approach was intended to supress the contribution from genes related to the general, non-specific pathways of the stress response to aggregation (i.e., quality control of proteostasis), thus highlighting those specific for RepA-WH1 cytotoxicity.