Yeast Prion Transformations

Prions are defined as infectious proteins that exist in regular non-prion or aggregated conformations. Prions play a crucial role in the epigenetic inheritance of traits in Saccharomyces cerevisiae. Among these, [PSI +] and [PIN +] are prion forms of Sup35p and Rnq1p, respectively. The presence of [PIN +] facilitates the induction of [PSI +], suggesting a key biochemical interaction between these two prions. This study aims to investigate their interaction through a series of genetic and fluorescence-based assays. We will eliminate [PIN +] in a 74D-694 strain using guanidine hydrochloride and transform both [PIN +] and [pin -] strains with p316CUP1-RNQ1sGFP and p316CUP1-NMsGFP plasmids. Fluorescence microscopy will be employed to analyze [PIN +] aggregation, and [PSI +] induction will be assessed by culturing transformed strains in SC-ura medium with CuSO₄. Further screening for [PSI +] isolates will be conducted on SC-ura-ade plates, followed by prion aggregation and colony color assays. This research will strengthen our understanding of prion interactions and their impact in proteinopathy such as Bovine spongiform encephalopathy and Alzheimer’s.