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Presenilin-1 differentially facilitates endoproteolysis of the β-amyloid precursor protein and Notch

Nature Cell Biology volume 2, pages 205–211 (2000)Cite this article

Abstract

Mutations in the presenilin-1 (PS1) gene are associated with Alzheimer’s disease and cause increased secretion of the neurotoxic amyloid-β peptide (Aβ). Critical intramembraneous aspartates at residues 257 and 385 are required for the function of PS1 protein. Here we investigate the biological function of a naturally occurring PS1 splice variant (PS1 Δexon 8), which lacks the critical aspartate 257. Cell lines that stably express PS1 Δexon 8 or a PS1 protein in which aspartate residue 257 is mutated secrete significant levels of Aβ, whereas Aβ generation is severely reduced in cells transfected with PS1 containing a mutation of aspartate 385. In contrast, endoproteolytic processing of Notch is almost completely inhibited in cell lines expressing any of the PS1 variants that lack one of the critical aspartates. These data indicate that PS1 may differentially facilitate γ-secretase-mediated generation of Aβ and endoproteolysis of Notch.

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Figure 1: Expression of PS1 Δexon 8, D257A and D385N inhibit presenilin-mediated endoproteolysis and differentially affect Aβ production.
Figure 2: Quantitative analysis of β-APP CTFs and Aβ in independent cell lines.
Figure 3: Expression of the active-site mutations as well as the splice variant of PS1 equally affect the accumulation of β-APP CTFs.
Figure 4: Differential inhibition of NICD and Aβ generation in cells stably expressing PS1 Δexon 8, D257A or D385N.
Figure 5: Subcellular distribution of β-APP in cells expressing endogenous presenilins or the indicated PS1 derivatives.
Figure 6: Cell-surface accumulation of β-APP and β-APP CTFs in cells expressing PS1 Δexon 8, D257A or D385N.
Figure 7: Differential distribution of Notch epitopes in cells expressing wild-type PS1, PS1 Δexon 8, PS1(D257A) or PS1(D385N) together with NotchΔE .

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Acknowledgements

This work was supported by a grant from the Deutsche Forschungsgemeinschaft (HA 1737/6-1) and the Boehringer Ingelheim K.G. We thank R. Kopan for the Notch cDNA construct, D. J. Selkoe for the PS1(D257A) cDNA, E. Koo for the monoclonal antibodies to β-APP, H-P. Hauri for the anti-giantin antibody, and P. Kahle for helpful discussions.

Correspondence and requests for materials should be addressed to C.H.

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Author notes

  1. Simone Keck & Miriam Baader

    Present address: Department of Molecular Biology, Central Institute of Mental Health, J5, 68159, Mannheim, Germany

  2. Anja Capell and Harald Steiner: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry Laboratory for Alzheimer’s Disease Research, Adolf Butenandt-Institute, Ludwig-Maximilians-University, Munich, 80336, Germany

    Anja Capell, Harald Steiner, Simone Keck, Miriam Baader & Christian Haass

  2. Boehringer Ingelheim KG, CNS Research, Ingelheim, 55216, Germany

    Helmut Romig

  3. Gene Center, Ludwig-Maximilians-University, Munich, 81377, Germany

    Melissa G. Grim & Ralf Baumeister

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Capell, A., Steiner, H., Romig, H. et al. Presenilin-1 differentially facilitates endoproteolysis of the β-amyloid precursor protein and Notch. Nat Cell Biol 2, 205–211 (2000). https://doi.org/10.1038/35008626

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