Figure 5.

PARK9 protects cells from elevated manganese levels. a) Examples of conditions used to identify the substrate specificity of YPK9. We identified ypk9Δ cells as being sensitive to manganese (Mn²⁺) relative to WT cells. b) The effect of various Mn²⁺ concentrations on ypk9Δ cells grown on rich (YPD) or synthetic (CSM) media. c) Expressing WT YPK9 in ypk9Δ cells is sufficient to rescue Mn²⁺ sensitivity but neither the PARK9 patient-based mutants nor the ATPase-dead mutant are able to rescue. Expressing YPK9 in WT yeast cells makes them more resistant to Mn²⁺ (compare top and bottom spottings). d) YFP- (top panel) and GFP-tagged (bottom panel) Ypk9 fusion proteins used for localization studies are functional because they are able to protect against Mn²⁺ sensitivity. e) ypk9Δ cells also exhibit sensitivity to Mn²⁺ when grown in liquid culture.