The gamma secretase-generated carboxyl-terminal domain of the amyloid precursor protein induces apoptosis via Tip60 in H4 cells.

The amyloid precursor protein (APP), a large glycoprotein highly expressed in neurons, is cleaved in its intramembranous domain by γ secretase to generate amyloid-β and a free carboxyl-terminal intracellular fragment (APP-CT), which has previously been suggested to interact with the adapter protein Fe65 and the histone acetyltransferase Tip60. An identical γ secretase activity mediates cleavage of Notch, releasing an intracellular signaling domain that translocates to the nucleus. We examined the effect of an ectopically expressed 58-amino acid APP-CT fragment (APP-C58) on human H4 neuroglioma cells. We demonstrate by confocal microscopy and fluorescence resonance energy transfer analysis that APP-C58 translocates to the nucleus and forms a complex in the nucleus with the Tip60, independent of interactions with Fe65. APP-C58 transfected H4 cells undergo apoptosis within 48–72 h, marked by nuclear blebbing, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, and blockade by a caspase inhibitor. When nuclear access of APP-C58 is prevented by fusing with a strong membrane-targeting farnesylation domain, apoptosis is blocked. APP-C58-induced apoptosis was markedly enhanced by co-transfection with wild type Tip60 and decreased by mutant Tip60 lacking histone acetyltransferase activity, suggesting that Tip60 mediates APP-CT-induced cell death. Thus, γ secretase cleavage of APP may contribute to Alzheimer's disease-related neurodegeneration in two ways: release of amyloid-β and liberation of a bioactive carboxyl-terminal domain from membrane-bound APP.

[1]  M. Fortini,et al.  Neurogenic phenotypes and altered Notch processing in Drosophila Presenilin mutants , 1999, Nature.

[2]  A. Prochiantz,et al.  A Short Cytoplasmic Domain of the Amyloid Precursor Protein Induces Apoptosis In Vitro and In Vivo , 2001, Molecular and Cellular Neuroscience.

[3]  P J McLean,et al.  Membrane association and protein conformation of alpha-synuclein in intact neurons. Effect of Parkinson's disease-linked mutations. , 2000, The Journal of biological chemistry.

[4]  B. Margolis,et al.  The phosphotyrosine interaction domains of X11 and FE65 bind to distinct sites on the YENPTY motif of amyloid precursor protein , 1996, Molecular and cellular biology.

[5]  D. Selkoe,et al.  The Intracellular Domain of the β-Amyloid Precursor Protein Is Stabilized by Fe65 and Translocates to the Nucleus in a Notch-like Manner* , 2001, The Journal of Biological Chemistry.

[6]  R. Barbour,et al.  Purification and cloning of amyloid precursor protein β-secretase from human brain , 1999, Nature.

[7]  S. Lincoln,et al.  A Loss of Function Mutation of Presenilin-2 Interferes with Amyloid β-Peptide Production and Notch Signaling* , 1999, The Journal of Biological Chemistry.

[8]  David G. Tew,et al.  Identification of a Novel Aspartic Protease (Asp 2) as β-Secretase , 1999, Molecular and Cellular Neuroscience.

[9]  J. Treanor,et al.  Beta-secretase cleavage of Alzheimer's amyloid precursor protein by the transmembrane aspartic protease BACE. , 1999, Science.

[10]  Marion Becker,et al.  Enhanced immunogenicity for CD8+ T cell induction and complete protective efficacy of malaria DNA vaccination by boosting with modified vaccinia virus Ankara , 1998, Nature Medicine.

[11]  Thomas C. Südhof,et al.  A Transcriptively Active Complex of APP with Fe65 and Histone Acetyltransferase Tip60 , 2001, Science.

[12]  B. Hyman,et al.  Aspartate Mutations in Presenilin and γ‐Secretase Inhibitors Both Impair Notch1 Proteolysis and Nuclear Translocation with Relative Preservation of Notch1 Signaling , 2000, Journal of neurochemistry.

[13]  Jun Qin,et al.  Involvement of the TIP60 Histone Acetylase Complex in DNA Repair and Apoptosis , 2000, Cell.

[14]  S. Pimplikar,et al.  The γ-secretase-cleaved C-terminal fragment of amyloid precursor protein mediates signaling to the nucleus , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  M. Citron.  Identifying Proteases That Cleave APP , 2000, Annals of the New York Academy of Sciences.

[16]  D. Selkoe,et al.  Cell Surface Presenilin-1 Participates in the γ-Secretase-like Proteolysis of Notch* , 1999, The Journal of Biological Chemistry.

[17]  B. Yankner,et al.  Proteolytic release and nuclear translocation of Notch-1 are induced by presenilin-1 and impaired by pathogenic presenilin-1 mutations. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[18]  B. de Strooper,et al.  A physiologic signaling role for the γ-secretase-derived intracellular fragment of APP , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[19]  S. Chandra,et al.  A second cytotoxic proteolytic peptide derived from amyloid β-protein precursor , 2000, Nature Medicine.

[20]  D. Price,et al.  Evidence that beta-amyloid protein in Alzheimer's disease is not derived by normal processing. , 1990, Science.

[21]  T. Oltersdorf,et al.  Cleavage of amyloid beta peptide during constitutive processing of its precursor. , 1990, Science.

[22]  Alfredo G. Tomasselli,et al.  Membrane-anchored aspartyl protease with Alzheimer's disease β-secretase activity , 1999, Nature.

[23]  B. Hyman,et al.  Demonstration by Fluorescence Resonance Energy Transfer of Two Sites of Interaction between the Low-Density Lipoprotein Receptor-Related Protein and the Amyloid Precursor Protein: Role of the Intracellular Adapter Protein Fe65 , 2001, The Journal of Neuroscience.

[24]  William J. Ray,et al.  A presenilin-1-dependent γ-secretase-like protease mediates release of Notch intracellular domain , 1999, Nature.

[25]  Klaus Fuchs,et al.  Presenilin‐dependent γ‐secretase processing of β‐amyloid precursor protein at a site corresponding to the S3 cleavage of Notch , 2001 .

[26]  Iva Greenwald,et al.  Presenilin is required for activity and nuclear access of Notch in Drosophila , 1999, Nature.

[27]  B. de Strooper,et al.  The amyloid precursor protein (APP)‐cytoplasmic fragment generated by γ‐secretase is rapidly degraded but distributes partially in a nuclear fraction of neurones in culture , 2001, Journal of neurochemistry.

[28]  G. Kotwal,et al.  Examination of the interactions of the amyloid precursor protein carboxyl terminus to intracellular protein: Possible role in apoptosis , 1999, Progress in Neuro-Psychopharmacology and Biological Psychiatry.

[29]  B. Hyman,et al.  Notch1 inhibits neurite outgrowth in postmitotic primary neurons , 1999, Neuroscience.