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A novel gene, Jpk, induces apoptosis in F9 murine teratocarcinoma cell through ROS generation

Kyoung-Ah Kong1, Sungdo Park, Hyoungwoo Park

  • 1Department of Anatomy, Embryology Laboratory, Brain Korea 21 Project for Medical Sciences, Yonsei University College of Medicine, Seoul 120-752, Korea.

Annals of the New York Academy of Sciences
|March 23, 2004
PubMed

Insights

A novel gene, Jopock (Jpk), induces apoptosis in eukaryotic cells by reducing mitochondrial membrane potential and increasing reactive oxygen species (ROS). This trans-acting factor is expressed during early gastrulation.

Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

Background:

  • A novel trans-acting factor, Jopock (Jpk), was identified through yeast hybridization, interacting with a murine Hoxa-7 regulatory element.
  • Jpk expression is detected at day 7.0 post coitum during early gastrulation in mice.
  • Previous studies indicated Jpk protein induces bacterial cell death.

Purpose of the Study:

  • To investigate the role and mechanism of Jpk in eukaryotic cell death.
  • To determine if Jpk induces apoptosis and its associated cellular events.

Main Methods:

  • Yeast hybridization for gene isolation.
  • Northern analysis for expression profiling.
  • F9 cell transfection and viability assays.
  • Flow cytometry using Annexin V/7-AAD, MitoTracker, and hydroethidine.
  • Reverse transcription PCR (RT-PCR) for gene expression analysis.

Main Results:

  • Jpk transfection in F9 cells led to cell death, with less than 50% survival after 72 hours.
  • Flow cytometry confirmed Jpk induces time-dependent apoptotic cell death.
  • Jpk reduced mitochondrial membrane potential and increased reactive oxygen species (ROS) production.
  • RT-PCR suggested Jpk regulates the Bcl family genes at the transcriptional level.

Conclusions:

  • Jpk is a potent inducer of apoptosis in eukaryotic cells.
  • Apoptosis induction by Jpk involves ROS generation and mitochondrial membrane potential reduction.
  • Jpk may regulate apoptosis through the Bcl family of proteins.

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