Relationship between alpha synuclein phosphorylation, proteasomal inhibition and cell death: relevance to Parkinson's disease pathogenesis.
Alpha synuclein can be phosphorylated at serine129 (P-S129), and the presence of highly phosphorylated alpha-synuclein in Lewy bodies suggests changes to its phosphorylation status has an important pathological role. We demonstrate that the kinase(s) responsible for alpha-synuclein S129 phosphorylation is constitutively active in SH-SY5Y cells and involves casein kinase 2 activity. Increased oxidative stress or proteasomal inhibition caused significant elevation of P-S129 alpha-synuclein levels. Under these conditions, similar increases in P-S129 alpha-synuclein were found in both sodium dodecyl sulphate lysates and Triton extracts indicating the phosphorylated protein was soluble and did not lead to aggregation. The rate of S129 phosphorylation was increased in response to proteasomal inhibition indicating a higher activity of the relevant kinase. Cells expressing the phosphorylation mimic, S129D alpha-synuclein increased cell death and enhanced sensitivity to epoxomycin exposure. Proteasomal inhibition markedly decreased S129D alpha-synuclein turnover suggesting proteasomal inhibition leads to the accumulation of P-S129 alpha-synuclein through an increase in the kinase activity and a decrease in protein turnover resulting in increased cell death. We conclude that S129 phosphorylation is toxic to dopaminergic cells and both the levels of S129 phosphorylated protein and its toxicity are increased with proteasomal inhibition emphasising the interdependence of these pathways in Parkinson's disease pathogenesis.
Chau
Kai-Yin
KY
University Department of Clinical Neurosciences, Institute of Neurology, University College London, London, UK.
Ching
Hey Long
HL
Schapira
Anthony H V
AH
Cooper
J Mark
JM
D016923:Cell Death / Q000502:physiology; D045744:Cell Line, Tumor; D006801:Humans; D010300:Parkinson Disease / Q000209:etiology* / Q000378:metabolism / Q000473:pathology*; D010766:Phosphorylation / Q000502:physiology; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D061988:Proteasome Inhibitors; D015854:Up-Regulation; D051844:alpha-Synuclein / Q000009:adverse effects / Q000096:biosynthesis / Q000378:metabolism*
D061988:Proteasome Inhibitors; D051844:alpha-Synuclein; D046988:Proteasome Endopeptidase Complex