Figure 4. Distinct regions of Rnq1 are important for each [RNQ+] variant.

(A) Summary of how disruption of the amyloidogenic regions by alanine mutations affects propagation of the [RNQ+] variants. Mutants are categorized as having no effect (white), mild effect (yellow), moderate effect (orange), or not propagating [RNQ+] (red) after successive passaging. These effects on [RNQ+] propagation were summarized from at least three independent experiments of each of the following assays: solubility assay, thermal stability, and SDD-AGE. See Figure S5 for more detail. If only one assay showed mild effects, e.g. SDD-AGE of s.d. low [RNQ+] cells harboring the A6 mutant, this was scored as having no effect in this summary table. (B) Mutation of the Rnq1 N-terminal domain shows [RNQ+] variant-dependent effects. Lysates from cells propagating s.d. high or m.d. high [RNQ+] and expressing wild-type (WT) Rnq1 or the N-terminal alanine mutants (A1–A5) were resolved by SDD-AGE, followed by western blot using an αRnq1 antibody. (C–E) Mutation of Rnq1 amyloidogenic regions differentially affects [RNQ+] variants. Yeast lysates propagating (C) s.d. low [RNQ+], (D) s.d. medium [RNQ+], or (E) s.d. very high [RNQ+] expressing WT Rnq1 or the set of alanine mutants (A1–A11) were analyzed by SDD-AGE and western blot with an αRnq1 antibody.