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Proteasome Inhibitors Prevent the Degradation of Familial Alzheimer’s Disease-Linked Presenilin 1 and Potentiate Aβ42 Recovery from Human Cells

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Abstract

Background

Several lines of evidence suggest that most of the early-onset forms of familial Alzheimer’s disease (FAD) are due to inherited mutations borne by a chromosome 14–encoded protein, presenilin 1 (PS1). This is likely related to an increased production of amyloid β-peptide (A β)42, one of the main components of the extracellular deposits called senile plaques that invade human cortical areas during the disease.

Materials and Methods

We set up stably transfected HEK293 cells overexpressing wild-type (wt) and various FAD-linked mutated PS1. By Western blot analysis, we examined the influence of specific proteasome inhibitors on PS1-like immunoreactivities. Furthermore, by means of metabolic labeling and immunoprecipitation with A β40 and A β42-directed specific antibodies, we assessed the effect of the inhibitors on the production of A βs by wt and mutated PS1-expressing cells transiently transfected with βAPP751.

Results

We show that two distinct proteasome inhibitors, Z-IE(Ot-Bu)A-Leucinal and lactacystin, increase in a time- and dose-dependent manner the immunoreactivities of both wt and mutated PS1. Furthermore, we demonstrate that PS1 is polyubiquitinated in these cells. Other inhibitors, ineffective on the proteasome, fail to protect wt and mutated PS1-like immunoreactivities. We also establish that the FAD-linked mutations of PS1 trigger a selective increased formation of Aβ42 as reflected by higher Aβ42 over total Aβ ratios when compared with wtPS1-expressing cells. Interestingly, this augmentation was further amplified by proteasome inhibitors in cells expressing mutated but not wtPS1.

Conclusion

Altogether, our data indicate that PS1 undergoes polyubiquitination in HEK293 cells and that the proteasome contributes to the degradation of wt and FAD-linked PS1, thereby directly influencing the Aβ production in human cells.

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Acknowledgements

We thank Drs. M. Savage and B. Greenberg (Cephalon, West Chester, PA) for generously providing us with 207 antibody and Dr. B. De Strooper (Center for Human Genetics, Leuven, Belgium) for the generous gift of the antibody (B14) directed toward the N-terminal part of PS1. We sincerely thank Dr. Sherwin Wilk for the generous gift of Z-IE(Ot-Bu)A-Leucinal and Z-L-Leucinal. We sincerely thank Drs. G. Thinakaran and S. Sisodia for generously providing us with αPS1Loop antibody and ΔE9-PS1 cDNA. We also thank J. Kervella for expert secretarial assistance, and Franck Aguila for the artwork. This work was supported by the Institut National de la Santé et de la Recherche Médicale and the Centre National de la Recherche Scientifique.

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Authors and Affiliations

  1. Institut de Pharmacologie Moléculaire et Cellulaire, UPR 411 du CNRS, 660 Route des lucioles, Sophia Antipolis, 06560, Valbonne, France

    Philippe Marambaud, Karine Ancolio, Elvira Lopez-Perez &  Frédéric Checler

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  1. Philippe Marambaud
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  2. Karine Ancolio
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  3. Elvira Lopez-Perez
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  4. Frédéric Checler
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Correspondence to Frédéric Checler.

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Communicated by P. Chambon.

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Marambaud, P., Ancolio, K., Lopez-Perez, E. et al. Proteasome Inhibitors Prevent the Degradation of Familial Alzheimer’s Disease-Linked Presenilin 1 and Potentiate Aβ42 Recovery from Human Cells. Mol Med 4, 147–157 (1998). https://doi.org/10.1007/BF03401912

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  • DOI: https://doi.org/10.1007/BF03401912

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