Enhanced α-syn toxicity in elongase null mutants.

<p>(A) The effect of WT α-syn expression on the growth of three deletion strains (<i>elo1</i>Δ, <i>elo2</i>Δ, and <i>elo3</i>Δ) and the wild-type BY4741 strain was evaluated. In panels (A, B), strains were transformed with pAG426GAL-WT (WT α-syn), pAG426GAL-A30P (A30P), pAG426GAL-A53T (A53T), pAG426GAL-E46K (E46K) or pAG426GAL (empty vector) and spotted in successive 10-fold serial dilutions on solid sucrose and solid galactose plates and incubated for 3 days at 30°C. Experiments were repeated three times with similar results. (B) The effect of the various α-syns (WT, A30P, A53T, or E46K) on the growth of the three <i>elo</i> deletion strains was also evaluated. (C) Western blot analysis of yeast cells expressing α-syn. Lysates were prepared from cultures grown for 6 h in inducing medium and then subjected to SDS–PAGE followed by western blot analysis. The cell-signaling polyclonal antibody against α-syn was used to visualize the three α-syns. The loading control was the yeast protein Pgk1p. Identical amounts of protein were loaded per well. Plasmids: see panel (A).