APP intracellular domain derived from amyloidogenic Î²- and Î³-secretase cleavage regulates neprilysin expression

Grimm, Marcus O. W.; Mett, Janine; Stahlmann, Christoph P; Grösgen, Sven; Haupenthal, Viola J.; Blümel, Tamara; Hundsdörfer, Benjamin; Zimmer, Valerie C.; Mylonas, Nadine; Tanila, Heikki; Müller, Ulrike; Grimm, Heike S.; Hartmann, Tobias

doi:10.3389/fnagi.2015.00077

Cited by 55 publications

(55 citation statements)

References 103 publications

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“…According to present literature 10–12 , a potential function of the APP-dependent nuclear aggregates is the modulation of gene expression in dependence of yet unknown stimuli. In order to test this hypothesis, transfected cells were fixed and stained with an anti-Histone3-K9ac antibody recognizing transcriptional active loci.…”

Section: Resultsmentioning

confidence: 99%

“…The presence of the histone acetyl transferase (TIP60) and the DNA helicase (BLM) in the complex points to a functional role in essential biological mechanisms such as gene expression, DNA replication/damage/repair or chromatin modification. Indeed, a variety of target genes like GSK3β, IDE, and APP have been proposed to be APP-CT dependently regulated 10–13 . In addition, some factors have been identified that modulate the subcellular localization of the APP-CT domain 13–17 as well as the molecules that are relevant for APP-CT-dependent DNA damage 18,19 .…”

Section: Introductionmentioning

confidence: 99%

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Amyloid precursor protein causes fusion of promyelocytic leukemia nuclear bodies in human hippocampal areas with high plaque load

Marks

Heinen

Bachmann

et al. 2020

Preprint

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The amyloid precursor protein (APP) is a type I transmembrane protein with unknown physiological function but potential impact in neurodegeneration. The current study demonstrates that APP signals to the nucleus causing the generation of aggregates comprising its adapter protein FE65 and the tumour suppressor proteins p53 and PML. The PML nuclear body generation, known to be of relevance in virus defence and cell division, is induced and fusion occurs over time depending on APP signalling. We further show that the nuclear aggregates of APP C-terminal (APP-CT) fragments together with PML and FE65 are present in the aged human brain but not in cerebral organoids differentiated from iPS cells.Notably, human Alzheimer's disease brains reveal a highly significant loss of these nuclear aggregates in areas with high plaque load compared to plaque-free areas of the same individual. Based on these results we conclude that APP-CT signalling to the nucleus takes DPBS (Gibco) and fixed in 4 % paraformaldehyde in PBS. Cover glasses were mounted with the Shandon™ Immu-Mount™ solution (Thermo Scientific) on glass slides and dried overnight at RT. For immunofluorescence staining, HEK293T cells were seeded in 8-well-µslide-ibi Treat (ibidi ® , Martinsried, Germany) and transfected using calcium phosphate transfection after 24 h. Cells were fixed with Roti ® -Histofix 4 % (4 % phosphate buffered formaldehyde solution; Roth, Karlsruhe, DE) for 20 min at 37 °C, and permeabilized and blocked with 5 % normal goat serum (NGS) in 0.3 % (w/v) Triton X-100/PBS for 30 min at RT. The cells were incubated with primary antibodies diluted in 1 % BSA/0.3 % Triton X-100/DPBS (mouse anti-HA (BioLegend, 901501; 1:1000), mouse anti-myc (NEB/Cell Signaling, 2276; 1:1500) o/n at 4 °C. For the secondary antibody staining and the cell staining, goat-anti-mouse AF568 (Invitrogen, A11004, 1:1000) was used, together with HCS CellMask™ Deep Red Stain (ThermoFisher Scientific, H32721, 1:5000), Hoechst33342 (10 mg/ml in H2O, Applichem, A0741, 1:1000) in DPBS (1 % BSA, 0.3 % Triton) and incubated for 1 h at RT.Immunoprecipitation HEK 293T cells were seeded in 10 cm dishes and co-transfection was performed 24 h after seeding. Whole cell extracts were prepared 24 h after transfection by scraping the cells from the dish with a cell scraper, washing the cell pellet in ice-cold PBS, extracting with 1 ml interaction buffer (50 mM Tris pH 8, 150 mM NaCl, 5 mM EDTA, 0.5% NP40, 1 mM DTT, 1 mM PMSF, 1x complete protease inhibitor cocktail), followed by sonication (15 s at 95 % amplitude) using a Sonopuls mini20 device (Bandelin, Berlin, Germany). The lysates were centrifuged (15,000 g, 15 min, 4 °C) and the supernatant was transferred to a new reaction tube. Input samples of the lysates were stored separately. Immunoprecipitation (IP) was carried out with the µMACS isolation kits for tagged proteins from Miltenyi Biotec (Bergisch-Gladbach, Germany). The eluates, as well as input samples of the lysates were subjected to SDS-PAGE and immunoblotting.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Amyloid precursor protein causes fusion of promyelocytic leukemia nuclear bodies in human hippocampal areas with high plaque load

Marks

Heinen

Bachmann

et al. 2020

Preprint

View full text Add to dashboard Cite

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“…Subsequently, tocopherol and tocotrienol (10 µM) were applied to the cells for 18 h followed by an additional treatment for 6 h with tocopherol and tocotrienol in combination with 0.5 µg/mL human synthetic Aβ40. To inhibit the activity of IDE [83], cells were additionally incubated with 10 µM human insulin (Sigma). Non-degraded human Aβ40 from cell culture supernatant was separated in SDS-PAGE and detected by Western blotting using the W02 antibody as described above.…”

Section: Methodsmentioning

confidence: 99%

Tocotrienol Affects Oxidative Stress, Cholesterol Homeostasis and the Amyloidogenic Pathway in Neuroblastoma Cells: Consequences for Alzheimer’s Disease

Grimm

Regner

Mett

et al. 2016

IJMS

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One of the characteristics of Alzheimer´s disease (AD) is an increased amyloid load and an enhanced level of reactive oxidative species (ROS). Vitamin E has known beneficial neuroprotective effects, and previously, some studies suggested that vitamin E is associated with a reduced risk of AD due to its antioxidative properties. However, epidemiological studies and nutritional approaches of vitamin E treatment are controversial. Here, we investigate the effect of α-tocotrienol, which belongs to the group of vitamin E, on AD-relevant processes in neuronal cell lines. In line with the literature, α-tocotrienol reduced the ROS level in SH-SY5Y cells. In the presence of tocotrienols, cholesterol and cholesterol esters, which have been shown to be risk factors in AD, were decreased. Besides the unambiguous positive effects of tocotrienol, amyloid-β (Aβ) levels were increased accompanied by an increase in the activity of enzymes responsible for Aβ production. Proteins and gene expression of the secretases and their components remained unchanged, whereas tocotrienol accelerates enzyme activity in cell-free assays. Besides enhanced Aβ production, tocotrienols inhibited Aβ degradation in neuro 2a (N2a)-cells. Our results might help to understand the controversial findings of vitamin E studies and demonstrate that besides the known positive neuroprotective properties, tocotrienols also have negative characteristics with respect to AD.

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“…A␤PP also influences the rate of protein synthesis [129], neurite outgrowth [130], and axonal pruning [131] as well as LPSmediated innate immune responses [132]. Signaling via AICD regulates genes including MEMBRANE METALLOENDOPEPTIDASE (MME, also known as neprilysin) [133] and TRANSTHYRETIN, TTR [134] that encode proteins involved in degradation and clearance of A␤ from the CNS, respectively. One or more of these functions may interact with PSEN holoprotein activities.…”

Section: What Then Of A␤pp A␤ and The A␤ 42 /A␤ 40 Ratio?mentioning

confidence: 99%

Evidence For and Against a Pathogenic Role of Reduced γ-Secretase Activity in Familial Alzheimer’s Disease

Jayne

Newman

Verdile

et al. 2016

JAD

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Abstract. The majority of mutations causing familial Alzheimer's disease (fAD) have been found in the gene PRESENILIN1 (PSEN1) with additional mutations in the related gene PRESENILIN2 (PSEN2). The best characterized function of PRESE-NILIN (PSEN) proteins is in ␥-secretase enzyme activity. One substrate of ␥-secretase is encoded by the gene AMYLOID BETA A4 PRECURSOR PROTEIN (A␤PP/APP) that is a fAD mutation locus. A␤PP is the source of the amyloid-␤ (A␤) peptide enriched in the brains of people with fAD or the more common, late onset, sporadic form of AD, sAD. These observations have resulted in a focus on ␥-secretase activity and A␤ as we attempt to understand the molecular basis of AD pathology. In this paper we briefly review some of the history of research on ␥-secretase in AD. We then discuss the main ideas regarding the role of ␥-secretase and the PSEN genes in this disease. We examine the significance of the "fAD mutation reading frame preservation rule" that applies to PSEN1 and PSEN2 (and A␤PP) and look at alternative roles for A␤PP and A␤ in fAD. We present a case for an alternative interpretation of published data on the role of ␥-secretase activity and fAD-associated mutations in AD pathology. Evidence supports a "PSEN holoprotein multimer hypothesis" where PSEN

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APP intracellular domain derived from amyloidogenic Î²- and Î³-secretase cleavage regulates neprilysin expression

Cited by 55 publications

References 103 publications

Amyloid precursor protein causes fusion of promyelocytic leukemia nuclear bodies in human hippocampal areas with high plaque load

Amyloid precursor protein causes fusion of promyelocytic leukemia nuclear bodies in human hippocampal areas with high plaque load

Tocotrienol Affects Oxidative Stress, Cholesterol Homeostasis and the Amyloidogenic Pathway in Neuroblastoma Cells: Consequences for Alzheimer’s Disease

Evidence For and Against a Pathogenic Role of Reduced γ-Secretase Activity in Familial Alzheimer’s Disease

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