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PAP gene transcription induced by cycloheximide in AR4-2J cells involves ADP-ribosylation

H Bödeker1, S Vasseur, N J Dusetti

  • 1U.315 INSERM, 46 boulevard de la Gaye, Marseille, F 13009, France.

Biochemical and Biophysical Research Communications
|October 29, 1998
PubMed

Insights

Cycloheximide increases pancreatic mRNA expression by directly regulating gene transcription. This process involves ADP-ribosylation, as indicated by inhibition experiments using poly(ADP-ribose) polymerase inhibitors.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Pancreatic acinar cell function is regulated by specific gene expression.
  • Cycloheximide is a known protein synthesis inhibitor used to study gene regulation.
  • The precise mechanisms of cycloheximide-induced gene expression in pancreatic cells are not fully understood.

Purpose of the Study:

  • To investigate the mechanism by which cycloheximide induces PAP mRNA expression in AR4-2J cells.
  • To determine whether cycloheximide affects PAP mRNA stability or directly induces transcription.
  • To explore the role of ADP-ribosylation in cycloheximide-mediated PAP gene upregulation.

Main Methods:

  • AR4-2J cells were treated with cycloheximide in a dose- and time-dependent manner.
  • Recombinant adenovirus carrying PAP cDNA was used to achieve high transcript levels.
  • Pharmacological inhibition of transcription followed by Northern blot analysis was performed.
  • Inhibition of poly(ADP-ribose) polymerase activity was assessed using specific chemical inhibitors.

Main Results:

  • Cycloheximide induced PAP mRNA expression in AR4-2J cells without affecting mRNA stability.
  • Inhibition of transcription did not alter the cycloheximide-induced increase in PAP mRNA levels over 4 hours.
  • Specific inhibitors of poly(ADP-ribose) polymerase (3-Aminobenzamide and 4-hydroxyquinazolone) significantly blocked PAP gene induction by cycloheximide.

Conclusions:

  • Cycloheximide induces PAP mRNA expression through direct regulation of gene transcription, not posttranscriptional stabilization.
  • ADP-ribosylation, mediated by poly(ADP-ribose) polymerase, is a critical component in the cycloheximide-induced upregulation of PAP gene expression.

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