title: Apoptotic cell death influences the signaling activity of the amyloid precursor protein through ShcA and Grb2 adaptor proteins in neuroblastoma SH-SY5Y cells.
abstract: The amyloid precursor protein (APP) is an ubiquitous receptor-like molecule involved in the pathogenesis of Alzheimer's disease (AD). APP and some of its C-terminal proteolytic fragments (CTFs) have been shown to be phosphorylated and to interact with cytosolic phosphotyrosine binding (PTB) domain containing proteins involved in cell signaling and vesicular transport. Among others, the interaction between tyrosine-phosphorylated CTFs and ShcA-Grb2 adaptors is highly enhanced in AD brain. Here we have identified in SH-SY5Y neuroblastoma cells an interaction between APP holoprotein and the adaptor Grb2. Upon activation of apoptotic cell death this interaction is rapidly degraded, APP is partially cleaved and the complex APP/Grb2 is replaced by a new complex between CTFs and ShcA that still involves Grb2. The formation of these complexes is regulated by beta-site APP-cleaving enzyme 1 and influences the phosphorylation of mitogen-activated protein kinase p44/42 extracellular signal-regulated kinase as well as the level of apoptotic death of the cells. These data suggest a dual role in cell signaling for APP and its CTFs in neuroblastoma cells, in a manner similar to that previously reported for other tyrosine kinase receptor, through a tightly regulated coupling with alternative intracellular adaptors to control the signaling of the cell.
authors.affiliation: Sezione di Farmacologia, Dipartimento di Oncologia, Biologia e Genetica, Università di Genova, e Dipartimento di Farmacologia e Neuroscienze IST c/o Centro di Biotecnologie Avanzate, Genova, Italy.
authors.forename: Valentina
authors.initials: V
authors.lastname: Venezia
authors.forename: Claudio
authors.initials: C
authors.lastname: Russo
authors.forename: Emanuela
authors.initials: E
authors.lastname: Repetto
authors.forename: Serena
authors.initials: S
authors.lastname: Salis
authors.forename: Virginia
authors.initials: V
authors.lastname: Dolcini
authors.forename: Francesca
authors.initials: F
authors.lastname: Genova
authors.forename: Mario
authors.initials: M
authors.lastname: Nizzari
authors.forename: Ulrike
authors.initials: U
authors.lastname: Mueller
authors.forename: Gennaro
authors.initials: G
authors.lastname: Schettini
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport / Q000276:immunology / Q000378:metabolism*; D053829:Amyloid Precursor Protein Secretases; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis / Q000502:physiology*; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors; D015153:Blotting, Western / Q000379:methods; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D010450:Endopeptidases; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005455:Fluorescent Antibody Technique / Q000379:methods; D051380:GRB2 Adaptor Protein; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D009447:Neuroblastoma; D010446:Peptide Fragments / Q000378:metabolism; D011233:Precipitin Tests / Q000379:methods; D011506:Proteins / Q000276:immunology / Q000378:metabolism*; D055765:Shc Signaling Adaptor Proteins; D015398:Signal Transduction / Q000502:physiology*; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D019311:Staurosporine / Q000494:pharmacology; D013997:Time Factors
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D033942:Adaptor Proteins, Vesicular Transport; D016564:Amyloid beta-Protein Precursor; D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D008164:Luminescent Proteins; D010446:Peptide Fragments; D011506:Proteins; C528024:SHC1 protein, human; D055765:Shc Signaling Adaptor Proteins; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D049452:Green Fluorescent Proteins; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D016282:Aspartic Acid Endopeptidases; C509280:BACE1 protein, human; D019311:Staurosporine