Tumor necrosis factor-alpha-elicited stimulation of gamma-secretase is mediated by c-Jun N-terminal kinase-dependent phosphorylation of presenilin and nicastrin.
Gamma-secretase is a multiprotein complex composed of presenilin (PS), nicastrin (NCT), Aph-1, and Pen-2, and it catalyzes the final proteolytic step in the processing of amyloid precursor protein to generate amyloid-beta. Our previous results showed that tumor necrosis factor-alpha (TNF-alpha) can potently stimulate gamma-secretase activity through a c-Jun N-terminal kinase ... (JNK)-dependent pathway. Here, we demonstrate that TNF-alpha triggers JNK-dependent serine/threonine phosphorylation of PS1 and NCT to stimulate gamma-secretase activity. Blocking of JNK activity with a potent JNK inhibitor (SP600125) reduces TNF-alpha-triggered phosphorylation of PS1 and NCT. Consistent with this, we show that activated JNKs can be copurified with gamma-secretase complexes and that active recombinant JNK2 can promote the phosphorylation of PS1 and NCT in vitro. Using site-directed mutagenesis and a synthetic peptide, we clearly show that the Ser(319)Thr(320) motif in PS1 is an important JNK phosphorylation site that is critical for the TNF-alpha-elicited regulation of gamma-secretase. This JNK phosphorylation of PS1 at Ser(319)Thr(320) enhances the stability of the PS1 C-terminal fragment that is necessary for gamma-secretase activity. Together, our findings strongly suggest that JNK is a critical intracellular mediator of TNF-alpha-elicited regulation of gamma-secretase and governs the pivotal step in the assembly of functional gamma-secretase.
Mesh Terms:
Amyloid Precursor Protein Secretases, Animals, CHO Cells, Cell Line, Cricetinae, Cricetulus, Enzyme Inhibitors, Gene Expression Regulation, Membrane Glycoproteins, Models, Biological, Mutation, Phosphorylation, Presenilins, Protein Binding, Protein Isoforms, Tumor Necrosis Factor-alpha
Amyloid Precursor Protein Secretases, Animals, CHO Cells, Cell Line, Cricetinae, Cricetulus, Enzyme Inhibitors, Gene Expression Regulation, Membrane Glycoproteins, Models, Biological, Mutation, Phosphorylation, Presenilins, Protein Binding, Protein Isoforms, Tumor Necrosis Factor-alpha
Mol. Biol. Cell
Date: Oct. 01, 2008
PubMed ID: 18667537
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