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Mol Med. 1999 March; 5(3): 160–168.
PMCID: PMC2230295

C-terminal maturation fragments of presenilin 1 and 2 control secretion of APP alpha and A beta by human cells and are degraded by proteasome.

Abstract

BACKGROUND: Most early-onset forms of Alzheimer's disease are due to missense mutations located on two homologous proteins named presenilin 1 and 2 (PS1 and PS2). Several lines of evidence indicate that PS1 and PS2 undergo various post-transcriptional events including endoproteolytic cleavages, giving rise to 28-30 kD N-terminal (NTF) and 18-20 kD C-terminal (CTF) fragments that accumulate in vivo. Whether the biological activity of presenilins is borne by the processed fragments or their holoprotein precursor remains in question. We have examined the putative control of beta APP maturation by CTF-PS1/PS2 and the catabolic process of the latter proteins by the multicatalytic complex, proteasome. MATERIALS AND METHODS: We transiently and stably transfected HEK293 cells with CTF-PS1 or CTF-PS2 cDNA. We examined these transfectants for their production of A beta 40, A beta 42, and APP alpha by immunoprecipitation using specific polyclonals. The effect of a series of proteases inhibitors on the immunoreactivity of CTF-PS1/PS2 was examined by Western blot. Finally, the influence of proteasome inhibitors on the generation of beta APP fragments by CTF-expressing cells was assessed by combined immunoprecipitation and densitometric analyses. RESULTS: We showed that transient and stable transfection of CTF-PS1 and CTF-PS2 cDNAs in human cells leads to increased secretion of APP alpha and A beta, the maturation products of beta APP. Furthermore, we demonstrated that two proteasome inhibitors, lactacystin and Z-IE(Ot-Bu)A-Leucinal, prevent the degradation of both CTFs. Accordingly, we established that proteasome inhibitors drastically potentiate the phenotypic increased production of APP alpha and A beta elicited by CTF-PS1/PS2. CONCLUSION: Our data establish that the C-terminal products of PS1 and PS2 maturation exhibit biological activity and in particular control beta APP maturation upstream to alpha-and beta/gamma-secretase cleavages. This function is directly controlled by the proteasome that modulates the intracellular concentration of CTFs.

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Selected References

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  • Tanzi RE, George-Hyslop PS, Gusella JF. Molecular genetics of Alzheimer disease amyloid. J Biol Chem. 1991 Nov 5;266(31):20579–20582. [PubMed]
  • Sherrington R, Rogaev EI, Liang Y, Rogaeva EA, Levesque G, Ikeda M, Chi H, Lin C, Li G, Holman K, et al. Cloning of a gene bearing missense mutations in early-onset familial Alzheimer's disease. Nature. 1995 Jun 29;375(6534):754–760. [PubMed]
  • Rogaev EI, Sherrington R, Rogaeva EA, Levesque G, Ikeda M, Liang Y, Chi H, Lin C, Holman K, Tsuda T, et al. Familial Alzheimer's disease in kindreds with missense mutations in a gene on chromosome 1 related to the Alzheimer's disease type 3 gene. Nature. 1995 Aug 31;376(6543):775–778. [PubMed]
  • Doan A, Thinakaran G, Borchelt DR, Slunt HH, Ratovitsky T, Podlisny M, Selkoe DJ, Seeger M, Gandy SE, Price DL, et al. Protein topology of presenilin 1. Neuron. 1996 Nov;17(5):1023–1030. [PubMed]
  • Lehmann S, Chiesa R, Harris DA. Evidence for a six-transmembrane domain structure of presenilin 1. J Biol Chem. 1997 May 2;272(18):12047–12051. [PubMed]
  • Takashima A, Sato M, Mercken M, Tanaka S, Kondo S, Honda T, Sato K, Murayama M, Noguchi K, Nakazato Y, et al. Localization of Alzheimer-associated presenilin 1 in transfected COS-7 cells. Biochem Biophys Res Commun. 1996 Oct 14;227(2):423–426. [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]
  • 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]
  • 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]
  • Mattson MP, Guo Q, Furukawa K, Pedersen WA. Presenilins, the endoplasmic reticulum, and neuronal apoptosis in Alzheimer's disease. J Neurochem. 1998 Jan;70(1):1–14. [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]
  • 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]
  • Baumann K, Paganetti PA, Sturchler-Pierrat C, Wong C, Hartmann H, Cescato R, Frey P, Yankner BA, Sommer B, Staufenbiel M. Distinct processing of endogenous and overexpressed recombinant presenilin 1. Neurobiol Aging. 1997 Mar-Apr;18(2):181–189. [PubMed]
  • Shirotani K, Takahashi K, Ozawa K, Kunishita T, Tabira T. Determination of a cleavage site of presenilin 2 protein in stably transfected SH-SY5Y human neuroblastoma cell lines. Biochem Biophys Res Commun. 1997 Nov 26;240(3):728–731. [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]
  • Loetscher H, Deuschle U, Brockhaus M, Reinhardt D, Nelboeck P, Mous J, Grünberg J, Haass C, Jacobsen H. Presenilins are processed by caspase-type proteases. J Biol Chem. 1997 Aug 15;272(33):20655–20659. [PubMed]
  • Kim TW, Pettingell WH, Jung YK, Kovacs DM, Tanzi RE. Alternative cleavage of Alzheimer-associated presenilins during apoptosis by a caspase-3 family protease. Science. 1997 Jul 18;277(5324):373–376. [PubMed]
  • Grunberg J, Walter J, Loetscher H, Deuschle U, Jacobsen H, Haass C. Alzheimer's disease associated presenilin-1 holoprotein and its 18-20 kDa C-terminal fragment are death substrates for proteases of the caspase family. Biochemistry. 1998 Feb 24;37(8):2263–2270. [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, Alves da Costa C, Ancolio K, Checler F. Alzheimer's disease-linked mutation of presenilin 2 (N141I-PS2) drastically lowers APPalpha secretion: control by the proteasome. Biochem Biophys Res Commun. 1998 Nov 9;252(1):134–138. [PubMed]
  • Marambaud P, Chevallier N, Ancolio K, Checler F. Post-transcriptional contribution of a cAMP-dependent pathway to the formation of alpha- and beta/gamma-secretases-derived products of beta APP maturation in human cells expressing wild-type and Swedish mutated beta APP. Mol Med. 1998 Nov;4(11):715–723. [PMC free article] [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]
  • 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]
  • Ida N, Hartmann T, Pantel J, Schröder J, Zerfass R, Förstl H, Sandbrink R, Masters CL, Beyreuther K. Analysis of heterogeneous A4 peptides in human cerebrospinal fluid and blood by a newly developed sensitive Western blot assay. J Biol Chem. 1996 Sep 13;271(37):22908–22914. [PubMed]
  • Fraser PE, Levesque G, Yu G, Mills LR, Thirlwell J, Frantseva M, Gandy SE, Seeger M, Carlen PL, St George-Hyslop P. Presenilin 1 is actively degraded by the 26S proteasome. Neurobiol Aging. 1998 Jan-Feb;19(1 Suppl):S19–S21. [PubMed]
  • Lee MK, Borchelt DR, Kim G, Thinakaran G, Slunt HH, Ratovitski T, Martin LJ, Kittur A, Gandy S, Levey AI, et al. Hyperaccumulation of FAD-linked presenilin 1 variants in vivo. Nat Med. 1997 Jul;3(7):756–760. [PubMed]
  • Vito P, Ghayur T, D'Adamio L. Generation of anti-apoptotic presenilin-2 polypeptides by alternative transcription, proteolysis, and caspase-3 cleavage. J Biol Chem. 1997 Nov 7;272(45):28315–28320. [PubMed]
  • Tomita T, Tokuhiro S, Hashimoto T, Aiba K, Saido TC, Maruyama K, Iwatsubo T. Molecular dissection of domains in mutant presenilin 2 that mediate overproduction of amyloidogenic forms of amyloid beta peptides. Inability of truncated forms of PS2 with familial Alzheimer's disease mutation to increase secretion of Abeta42. J Biol Chem. 1998 Aug 14;273(33):21153–21160. [PubMed]

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