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. Author manuscript; available in PMC: 2018 Apr 1.
Published in final edited form as: Mol Microbiol. 2017 Jan 26;104(1):105–124. doi: 10.1111/mmi.13616

Figure 3. The effect of altering Hsp104 and Sse1 activity on [SWI+] propagation.

Figure 3

(A) FLO8-repaired (FLO8) and unrepaired (flo8) BY4741 [SWI+] strains were transformed with p415F139-URA3, and streaked on SC selective plates (SC) with (+) or without (−) 5 mM GdnHCl and then cultured in SC-leu which was followed by growth on the indicated SC selective plates. Shown was a result (3 days post-spotting) of at least 3 independent experiments. (B, C, and D) As described in the Experimental Procedures, the influences of Hsp104 overproduction (HSP104), Hsp104 inactivation (GdnHCl), and Sse1 overproduction (SSE1) on [SWI+] propagation in 74D-694 (B) and BY4741 (C and D) backgrounds were investigated. vector, an empty vector control; SC, SC selective plate; 5FOA, SC selective plate with 5-FOA; Raf, raffinose plate; n, total colonies assayed. Note: the plasmid reporter PF139-URA3 was used for panel B and C, but chromosomal reporter of PFLO1-URA3 for panel D. Prion curing was summarized in the plot (right panel) based on the growth phenotypes (left). (E) Effect of Sse1 or Hsp104 overproduction on [SWI+] curing by GdnHCl was assayed as described in the Experimental Procedures. The remaining [SWI+] was assayed after growing cells in SC+GdnHCl medium for the indicated time while overproducing Hsp104 or Sse1. T-test was used to estimate the significance of the differences (N.S., not significant; **, P<0.01; ***, P<0.001). n, colonies assayed.