The β-amyloid peptide is generated by the proteolysis of the amyloid precursor protein (APP) by the action of β- and γ-secretase. The mechanisms underlying this process are poorly understood. Using a cell-based reporter gene assay we analysed the possible signals and pathways that could be involved in APP cleavage. We used the stable cell line HeLa AG that expresses the human APP(695) fused with the yeast transcription factor Gal4. This fusion protein is normally translocated into the plasma membrane and after APP-Gal4 cleavage, the AICD-Gal4 fragment released can activate the transcription of a luciferase reporter gene. Through this reporter system, we demonstrated that Ras GTPase, but not Ral and Rap, could promote APP-Gal4 cleavage. In addition HeLa AG cells stimulated with EGF or PDGF or overexpressing EGFR exhibit increased APP proteolysis in a Ras-dependent way. This process is also dependent on γ-secretase activity, being abolished by the γ-secretase inhibitor DAPT.

Amigoni, L., Ceriani, M., Belotti, F., Minopoli, G., Martegani, E. (2011). Activation of amyloid precursor protein processing by growth factors is dependent on Ras GTPase activity. NEUROCHEMICAL RESEARCH, 36(3), 392-398 [10.1007/s11064-010-0343-8].

Activation of amyloid precursor protein processing by growth factors is dependent on Ras GTPase activity

AMIGONI, LOREDANA;CERIANI, MICHELA;BELOTTI, FIORELLA;MARTEGANI, ENZO
2011

Abstract

The β-amyloid peptide is generated by the proteolysis of the amyloid precursor protein (APP) by the action of β- and γ-secretase. The mechanisms underlying this process are poorly understood. Using a cell-based reporter gene assay we analysed the possible signals and pathways that could be involved in APP cleavage. We used the stable cell line HeLa AG that expresses the human APP(695) fused with the yeast transcription factor Gal4. This fusion protein is normally translocated into the plasma membrane and after APP-Gal4 cleavage, the AICD-Gal4 fragment released can activate the transcription of a luciferase reporter gene. Through this reporter system, we demonstrated that Ras GTPase, but not Ral and Rap, could promote APP-Gal4 cleavage. In addition HeLa AG cells stimulated with EGF or PDGF or overexpressing EGFR exhibit increased APP proteolysis in a Ras-dependent way. This process is also dependent on γ-secretase activity, being abolished by the γ-secretase inhibitor DAPT.
Articolo in rivista - Articolo scientifico
Peptide Fragments; ras Proteins; Receptor, Epidermal Growth Factor; HeLa Cells; Enzyme Activation; Humans; Genes, Reporter; Recombinant Fusion Proteins; Intercellular Signaling Peptides and Proteins; Amyloid beta-Protein Precursor
English
2011
36
3
392
398
none
Amigoni, L., Ceriani, M., Belotti, F., Minopoli, G., Martegani, E. (2011). Activation of amyloid precursor protein processing by growth factors is dependent on Ras GTPase activity. NEUROCHEMICAL RESEARCH, 36(3), 392-398 [10.1007/s11064-010-0343-8].
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/10281/49025
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