FIGURE 7:

Non-PIKK phosphomimetic substitution decreases FUS toxicity and prion-like aggregation in yeast. (A) Schematic of the various phosphomimetic constructs used in the lab. Solid gray circles indicate PIKK consensus sites and light gray circles indicate non-PIKK sites. The constructs are in red to indicate their use in subsequent experiments. The black-highlighted axis indicates the FUS fragment inserted into Sup35 and used in Panel E. (B) Phosphomimetic substitution in the prion-like domain rescues FUS toxicity in yeast. (C) Ectopic expression of FUS 4Ev3 and 4Ev4 analyzed by structured illumination microscopy. Cells were probed with anti-FUS. (D) Quantification of FUS signal in punctate structures compared with total FUS expression; error bars represent 95% CI. Figure data analyzed using a Student t-test (n = 9) (E) Sup35-FUS or Sup35-FUS 4E were expressed in yeast on SC or SC-ade media. Sup35NM was added to promote prion formation under both conditions.