Skip to main content
. Author manuscript; available in PMC: 2009 Sep 1.
Published in final edited form as: Nat Genet. 2009 Feb 1;41(3):308–315. doi: 10.1038/ng.300

Figure 4.

Figure 4

Ypk9 is localized to the vacuole in yeast and PARK9 patient-based mutations affect its ability to rescue α–syn toxicity. a) Fluorescence microscopy to visualize Ypk9 subcellular localization. A chromosomally tagged YFP fusion (YPK9-YFP) localizes to the vacuolar membrane, as does WT GFP-YPK9 expressed from the constitutive GPD promoter. PARK9 patient-based mutations 23 alter YPK9 localization but the ATPase-dead mutant (D781N) does not. GFP-tagged human ATP13A2 also localizes to the vacuole in yeast cells. b) Spotting assays with WT or 〈–syn-expressing cells. WT YPK9 overexpression suppresses 〈–syn toxicity but the two PARK9 patient-based mutant YPK9 proteins as well as the ATPase-dead mutant do not. Expressing mutant YPK9 in WT cells does not inhibit growth, supporting the idea that these are loss-of-function and not dominant negative mutations.