Loss of function mutations in the gene encoding Omi/HtrA2 in Parkinson's diseaseRecently targeted disruption of Omi/HtrA2 has been found to cause neurodegeneration and a parkinsonian phenotype in mice. Using a candidate gene approach, we performed a mutation screening of the Omi/HtrA2 gene in German Parkinson's disease (PD) patients. In four patients, we identified a novel heterozygous G399S mutation, which was absent in healthy controls. Moreover, we identified a novel A141S polymorphism that was associated with PD (P<0.05). Both mutations resulted in defective activation of the protease activity of Omi/HtrA2. Immunohistochemistry and functional analysis in stably transfected cells revealed that S399 mutant Omi/HtrA2 and to a lesser extent, the risk allele of the A141S polymorphism induced mitochondrial dysfunction associated with altered mitochondrial morphology. Cells overexpressing S399 mutant Omi/HtrA2 were more susceptible to stress-induced cell death than wild-type. On the basis of functional genomics, our results provide a novel link between mitochondrial dysfunction and neurodegeneration in PD.
The proteasomal subunit S6 ATPase is a novel synphilin‐1 interacting protein—implications for Parkinson's diseaseABSTRACT Synphilin‐1 is linked to Parkinson's disease (PD), based on its role as an alpha‐synuclein (PARK1)‐interacting protein and substrate of the ubiquitin E3 ligase Parkin (PARK2) and because of its presence in Lewy bodies (LB) in brains of PD patients. We found that overexpression of synphilin‐1 in cells leads to the formation of ubiquitinated cytoplasmic inclusions supporting a derangement of the ubiquitin‐proteasome system in PD. We report here a novel specific interaction of synphilin‐1 with the regulatory proteasomal protein S6 ATPase (tbp7). Functional characterization of this interaction on a cellular level revealed colocalization of S6 and synphilin‐1 in aggresome‐like intracytoplasmic inclusions. Overexpression of synphilin‐1 and S6 in cells caused reduced proteasomal activity associated with a significant increase in inclusion formation compared to cells expressing syn‐philin‐1 alone. Steady‐state levels of synphilin‐1 in cells were not altered after cotransfection of S6 and colocal‐ization of synphilin‐1‐positive inclusions with lysosomal markers suggests the presence of an alternative lysosomal degradation pathway. Subsequent immunohistochemical studies in brains of PD patients identified S6 ATPase as a component of LB. This is the first study investigating the physiological role of synphilin‐1 in the ubiquitin protea‐some system. Our data suggest a direct interaction of synphilin‐1 with the regulatory complex of the protea‐some modulating proteasomal function.—Marx F. P., Soehn, A. S., Berg, D., Melle, C., Schiesling, C., Lang, M., Kautzmann, S., Strauss, K. M., Franck, T., Engelender, S., Pahnke, J., Dawson, S., von Eggeling F., Schulz, J. B., Riess, O., Krüger R. The proteasomal subunit S6 ATPase is a novel synphilin‐1 interacting protein—implications for Parkinson's disease. FASEB J. 21, 1759–1767 (2007)