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Quantitative Measurement of γ-Secretase-mediated Amyloid Precursor Protein and Notch Cleavage in Cell-based Luciferase Reporter Assay Platforms
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Gene expression profiling in cells with enhanced gamma-secretase activity

Alexandra I Magold1, Matthias Cacquevel, Patrick C Fraering

  • 1Brain Mind Institute, School of Life Sciences, Swiss Federal Institute of Technology (EPFL), Lausanne, Switzerland.

Plos One
|September 19, 2009
PubMed
Abstract

Insights

Gamma-secretase activity impacts gene transcription, affecting pathways relevant to cancer and Alzheimer's disease. This study identified 21 key genes, including TERA, altered by this enzyme.

Area of Science:

  • Molecular Biology
  • Genomics
  • Cellular Biology

Background:

  • Gamma-secretase processes type-I membrane proteins, releasing intracellular domains (ICDs) that modulate gene transcription.
  • Gamma-secretase is a therapeutic target for cancer and Alzheimer's disease.
  • Understanding gamma-secretase's transcriptional targets is crucial for disease research.

Purpose of the Study:

  • To identify genes and molecular functions transcriptionally regulated by gamma-secretase activity.
  • To investigate the role of gamma-secretase in cellular processes relevant to disease.

Main Methods:

  • Comparative transcriptomic analysis of Chinese hamster ovary (CHO) cells with altered gamma-secretase activity using cDNA microarray.
  • Functional clustering of identified genes using FatiGO.
  • Validation of key genes by real-time PCR.

Main Results:

  • Analysis identified 1,981 genes transcriptionally affected by gamma-secretase activity.
  • Twenty-one genes were validated, including decreased transcription of PTPRG and AMN1, and increased transcription of UPP1, relevant to cell cycle.
  • Significant alterations in Wnt pathway members (WNT3A, DVL3) and TERA (a gene of unknown function) were observed.
  • TERA transcription was also altered in Alzheimer's disease brain cortices.

Conclusions:

  • Gamma-secretase activity influences transcription of diverse molecular functions and specific genes.
  • The 21 validated genes provide insights into gamma-secretase's role in cancer and Alzheimer's disease.
  • TERA represents a novel gene potentially linked to Alzheimer's pathology.

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