Hsp90 interaction affects PINK1 subcellular distribution in Hela cells. A) Subcellular distribution of WT and L347P PINK1 in Hela cells is not noticeably different. C = cytosolic fraction; M = mitochondrial fraction. B) Subcellular distribution of mito-Δ151 PINK1 shows the loss of cytosolic PINK1 when cells are treated with 1 μM 17-AAG, a Hsp90 inhibitor, for 4 hours. C) mito-L347P PINK1 localizes to cytosol and mitochondria. D) Mito-L347P PINK1 accumulates more in the mitochondria than mito-Δ151 PINK1, with little change to the cytosol distribution. E) Quantification from three independent experiments showing more PINK1 in mitochondrial fraction with L347P mutation. P = 0.025. Increase in PINK1 in the cytosolic fraction with L347P is not statistically significant. Mean ± SEM, n = 3. Statistical significance was calculated with ANOVA and Fisher's PLSD post-hoc test. F) L347P mutation reduces PINK1 interaction with Hsp90 by co-immunoprecipitation. GFP does not co-immunoprecipitate with Hsp90. G) Summary diagram depicting PINK1 protein structures and the role of each component in PINK1 topology and subcellular distribution. See Discussion for more details.
Lin and Kang BMC Cell Biology 2010 11:90 doi:10.1186/1471-2121-11-90