Search tips
Search criteria 


Logo of molmedLink to Publisher's site
Mol Med. 1998 March; 4(3): 147–157.
PMCID: PMC2230353

Proteasome inhibitors prevent the degradation of familial Alzheimer's disease-linked presenilin 1 and potentiate A beta 42 recovery from human cells.


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 beta-peptide (A beta) 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 beta 40 and A beta 42-directed specific antibodies, we assessed the effect of the inhibitors on the production of A beta s by wt and mutated PS1-expressing cells transiently transfected with beta APP751. 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 A beta 42 as reflected by higher A beta 42 over total A beta 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 A beta production in human cells.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Lemere CA, Lopera F, Kosik KS, Lendon CL, Ossa J, Saido TC, Yamaguchi H, Ruiz A, Martinez A, Madrigal L, et al. The E280A presenilin 1 Alzheimer mutation produces increased A beta 42 deposition and severe cerebellar pathology. Nat Med. 1996 Oct;2(10):1146–1150. [PubMed]
  • Mann DM, Iwatsubo T, Cairns NJ, Lantos PL, Nochlin D, Sumi SM, Bird TD, Poorkaj P, Hardy J, Hutton M, et al. Amyloid beta protein (Abeta) deposition in chromosome 14-linked Alzheimer's disease: predominance of Abeta42(43). Ann Neurol. 1996 Aug;40(2):149–156. [PubMed]
  • Mann DM, Iwatsubo T, Nochlin D, Sumi SM, Levy-Lahad E, Bird TD. Amyloid (Abeta) deposition in chromosome 1-linked Alzheimer's disease: the Volga German families. Ann Neurol. 1997 Jan;41(1):52–57. [PubMed]
  • Borchelt DR, Thinakaran G, Eckman CB, Lee MK, Davenport F, Ratovitsky T, Prada CM, Kim G, Seekins S, Yager D, et al. Familial Alzheimer's disease-linked presenilin 1 variants elevate Abeta1-42/1-40 ratio in vitro and in vivo. Neuron. 1996 Nov;17(5):1005–1013. [PubMed]
  • Gearing M, Tigges J, Mori H, Mirra SS. A beta40 is a major form of beta-amyloid in nonhuman primates. Neurobiol Aging. 1996 Nov-Dec;17(6):903–908. [PubMed]
  • Xia W, Zhang J, Kholodenko D, Citron M, Podlisny MB, Teplow DB, Haass C, Seubert P, Koo EH, Selkoe DJ. Enhanced production and oligomerization of the 42-residue amyloid beta-protein by Chinese hamster ovary cells stably expressing mutant presenilins. J Biol Chem. 1997 Mar 21;272(12):7977–7982. [PubMed]
  • Tomita T, Maruyama K, Saido TC, Kume H, Shinozaki K, Tokuhiro S, Capell A, Walter J, Grünberg J, Haass C, et al. The presenilin 2 mutation (N141I) linked to familial Alzheimer disease (Volga German families) increases the secretion of amyloid beta protein ending at the 42nd (or 43rd) residue. Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):2025–2030. [PubMed]
  • Duff K, Eckman C, Zehr C, Yu X, Prada CM, Perez-tur J, Hutton M, Buee L, Harigaya Y, Yager D, et al. Increased amyloid-beta42(43) in brains of mice expressing mutant presenilin 1. Nature. 1996 Oct 24;383(6602):710–713. [PubMed]
  • Ancolio K, Marambaud P, Dauch P, Checler F. Alpha-secretase-derived product of beta-amyloid precursor protein is decreased by presenilin 1 mutations linked to familial Alzheimer's disease. J Neurochem. 1997 Dec;69(6):2494–2499. [PubMed]
  • Kovacs DM, Fausett HJ, Page KJ, Kim TW, Moir RD, Merriam DE, Hollister RD, Hallmark OG, Mancini R, Felsenstein KM, et al. Alzheimer-associated presenilins 1 and 2: neuronal expression in brain and localization to intracellular membranes in mammalian cells. Nat Med. 1996 Feb;2(2):224–229. [PubMed]
  • Cook DG, Sung JC, Golde TE, Felsenstein KM, Wojczyk BS, Tanzi RE, Trojanowski JQ, Lee VM, Doms RW. Expression and analysis of presenilin 1 in a human neuronal system: localization in cell bodies and dendrites. Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):9223–9228. [PubMed]
  • Walter J, Capell A, Grünberg J, Pesold B, Schindzielorz A, Prior R, Podlisny MB, Fraser P, Hyslop PS, Selkoe DJ, et al. The Alzheimer's disease-associated presenilins are differentially phosphorylated proteins located predominantly within the endoplasmic reticulum. Mol Med. 1996 Nov;2(6):673–691. [PMC free article] [PubMed]
  • Dewji NN, Singer SJ. Specific transcellular binding between membrane proteins crucial to Alzheimer disease. Proc Natl Acad Sci U S A. 1996 Oct 29;93(22):12575–12580. [PubMed]
  • Weidemann A, Paliga K, Dürrwang U, Czech C, Evin G, Masters CL, Beyreuther K. Formation of stable complexes between two Alzheimer's disease gene products: presenilin-2 and beta-amyloid precursor protein. Nat Med. 1997 Mar;3(3):328–332. [PubMed]
  • Marambaud P, Chevallier N, Barelli H, Wilk S, Checler F. Proteasome contributes to the alpha-secretase pathway of amyloid precursor protein in human cells. J Neurochem. 1997 Feb;68(2):698–703. [PubMed]
  • Marambaud P, Lopez-Perez E, Wilk S, Checler F. Constitutive and protein kinase C-regulated secretory cleavage of Alzheimer's beta-amyloid precursor protein: different control of early and late events by the proteasome. J Neurochem. 1997 Dec;69(6):2500–2505. [PubMed]
  • Marambaud P, Wilk S, Checler F. Protein kinase A phosphorylation of the proteasome: a contribution to the alpha-secretase pathway in human cells. J Neurochem. 1996 Dec;67(6):2616–2619. [PubMed]
  • Thinakaran G, Borchelt DR, Lee MK, Slunt HH, Spitzer L, Kim G, Ratovitsky T, Davenport F, Nordstedt C, Seeger M, et al. Endoproteolysis of presenilin 1 and accumulation of processed derivatives in vivo. Neuron. 1996 Jul;17(1):181–190. [PubMed]
  • Chevallier N, Jiracek J, Vincent B, Baur CP, Spillantini MG, Goedert M, Dive V, Checler F. Examination of the role of endopeptidase in A beta secretion by human transfected cells. Br J Pharmacol. 1997 Jun;121(3):556–562. [PMC free article] [PubMed]
  • Barelli H, Lebeau A, Vizzavona J, Delaere P, Chevallier N, Drouot C, Marambaud P, Ancolio K, Buxbaum JD, Khorkova O, et al. Characterization of new polyclonal antibodies specific for 40 and 42 amino acid-long amyloid beta peptides: their use to examine the cell biology of presenilins and the immunohistochemistry of sporadic Alzheimer's disease and cerebral amyloid angiopathy cases. Mol Med. 1997 Oct;3(10):695–707. [PMC free article] [PubMed]
  • Fenteany G, Standaert RF, Lane WS, Choi S, Corey EJ, Schreiber SL. Inhibition of proteasome activities and subunit-specific amino-terminal threonine modification by lactacystin. Science. 1995 May 5;268(5211):726–731. [PubMed]
  • Ward RV, Davis JB, Gray CW, Barton AJ, Bresciani LG, Caivano M, Murphy VF, Duff K, Hutton M, Hardy J, et al. Presenilin-1 is processed into two major cleavage products in neuronal cell lines. Neurodegeneration. 1996 Dec;5(4):293–298. [PubMed]
  • De Strooper B, Beullens M, Contreras B, Levesque L, Craessaerts K, Cordell B, Moechars D, Bollen M, Fraser P, George-Hyslop PS, et al. Phosphorylation, subcellular localization, and membrane orientation of the Alzheimer's disease-associated presenilins. J Biol Chem. 1997 Feb 7;272(6):3590–3598. [PubMed]
  • Seeger M, Nordstedt C, Petanceska S, Kovacs DM, Gouras GK, Hahne S, Fraser P, Levesque L, Czernik AJ, George-Hyslop PS, et al. Evidence for phosphorylation and oligomeric assembly of presenilin 1. Proc Natl Acad Sci U S A. 1997 May 13;94(10):5090–5094. [PubMed]
  • Walter J, Grünberg J, Capell A, Pesold B, Schindzielorz A, Citron M, Mendla K, George-Hyslop PS, Multhaup G, Selkoe DJ, et al. Proteolytic processing of the Alzheimer disease-associated presenilin-1 generates an in vivo substrate for protein kinase C. Proc Natl Acad Sci U S A. 1997 May 13;94(10):5349–5354. [PubMed]
  • Kim TW, Pettingell WH, Hallmark OG, Moir RD, Wasco W, Tanzi RE. Endoproteolytic cleavage and proteasomal degradation of presenilin 2 in transfected cells. J Biol Chem. 1997 Apr 25;272(17):11006–11010. [PubMed]
  • Mengual E, Arizti P, Rodrigo J, Giménez-Amaya JM, Castaño JG. Immunohistochemical distribution and electron microscopic subcellular localization of the proteasome in the rat CNS. J Neurosci. 1996 Oct 15;16(20):6331–6341. [PubMed]
  • Palmer A, Rivett AJ, Thomson S, Hendil KB, Butcher GW, Fuertes G, Knecht E. Subpopulations of proteasomes in rat liver nuclei, microsomes and cytosol. Biochem J. 1996 Jun 1;316(Pt 2):401–407. [PubMed]
  • Oda K, Ikehara Y, Omura S. Lactacystin, an inhibitor of the proteasome, blocks the degradation of a mutant precursor of glycosylphosphatidylinositol-linked protein in a pre-Golgi compartment. Biochem Biophys Res Commun. 1996 Feb 27;219(3):800–805. [PubMed]
  • Werner ED, Brodsky JL, McCracken AA. Proteasome-dependent endoplasmic reticulum-associated protein degradation: an unconventional route to a familiar fate. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13797–13801. [PubMed]
  • Fuller SJ, Storey E, Li QX, Smith AI, Beyreuther K, Masters CL. Intracellular production of beta A4 amyloid of Alzheimer's disease: modulation by phosphoramidon and lack of coupling to the secretion of the amyloid precursor protein. Biochemistry. 1995 Jun 27;34(25):8091–8098. [PubMed]
  • Haass C. Presenile because of presenilin: the presenilin genes and early onset Alzheimer's disease. Curr Opin Neurol. 1996 Aug;9(4):254–259. [PubMed]
  • Hardy J, Allsop D. Amyloid deposition as the central event in the aetiology of Alzheimer's disease. Trends Pharmacol Sci. 1991 Oct;12(10):383–388. [PubMed]
  • Checler F. Processing of the beta-amyloid precursor protein and its regulation in Alzheimer's disease. J Neurochem. 1995 Oct;65(4):1431–1444. [PubMed]
  • Rivett AJ. The multicatalytic proteinase. Multiple proteolytic activities. J Biol Chem. 1989 Jul 25;264(21):12215–12219. [PubMed]
  • Rechsteiner M, Hoffman L, Dubiel W. The multicatalytic and 26 S proteases. J Biol Chem. 1993 Mar 25;268(9):6065–6068. [PubMed]

Articles from Molecular Medicine are provided here courtesy of The Feinstein Institute for Medical Research at North Shore LIJ