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Journal article
An AICD-based functional screen to identify APP metabolism regulators
Molecular neurodegeneration, v 2(1), 15
01 Jan 2007
PMID: 17718916
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
Background: A central event in Alzheimer's disease (AD) is the regulated intramembraneous proteolysis of the beta-amyloid precursor protein (APP), to generate the beta-amyloid (A beta) peptide and the APP intracellular domain (AICD). A beta is the major component of amyloid plaques and AICD displays transcriptional activation properties. We have taken advantage of AICD transactivation properties to develop a genetic screen to identify regulators of APP metabolism. This screen relies on an APP-Gal4 fusion protein, which upon normal proteolysis, produces AICD-Gal4. Production of AICD-Gal4 induces Gal4-UAS driven luciferase expression. Therefore, when regulators of APP metabolism are modulated, luciferase expression is altered.
Results: To validate this experimental approach we modulated alpha-, beta-, and gamma-secretase levels and activities. Changes in AICD-Gal4 levels as measured by Western blot analysis were strongly and significantly correlated to the observed changes in AICD-Gal4 mediated luciferase activity. To determine if a known regulator of APP trafficking/maturation and Presenilin1 endoproteolysis could be detected using the AICD-Gal4 mediated luciferase assay, we knocked-down Ubiquilin 1 and observed decreased luciferase activity. We confirmed that Ubiquilin 1 modulated AICD-Gal4 levels by Western blot analysis and also observed that Ubiquilin 1 modulated total APP levels, the ratio of mature to immature APP, as well as PS1 endoproteolysis.
Conclusion: Taken together, we have shown that this screen can identify known APP metabolism regulators that control proteolysis, intracellular trafficking, maturation and levels of APP and its proteolytic products. We demonstrate for the first time that Ubiquilin 1 regulates APP metabolism in the human neuroblastoma cell line, SH-SY5Y.
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Details
- Title
- An AICD-based functional screen to identify APP metabolism regulators
- Creators
- Can Zhang - Drexel UniversityPreeti J. Khandelwal - Drexel UniversityRanjita Chakraborty - Drexel UniversityTrinna L. Cuellar - Drexel UniversitySrikant Sarangi - Drexel UniversityShyam A. Patel - Drexel UniversityChristopher P. Cosentino - Drexel UniversityMichael O'Connor - Drexel UniversityJeremy C. Lee - Drexel UniversityRudolph E. Tanzi - Massachusetts General HospitalAleister J. Saunders - Drexel University
- Publication Details
- Molecular neurodegeneration, v 2(1), 15
- Publisher
- Springer Nature
- Number of pages
- 19
- Grant note
- R01NS057295 / NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE; United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Neurological Disorders & Stroke (NINDS) Drexel University NINDS (AJS); United States Department of Health & Human Services; National Institutes of Health (NIH) - USA; NIH National Institute of Neurological Disorders & Stroke (NINDS) Commonwealth of PA
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biodiversity, Earth, and Environmental Science (BEES); Biology
- Web of Science ID
- WOS:000206907200015
- Scopus ID
- 2-s2.0-36048978614
- Other Identifier
- 991019169410704721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Neurosciences