PC12 cells regulate inducible cyclic AMP (cAMP) element repressor expression to differentially control cAMP response element-dependent transcription in response to nerve growth factor and cAMP

J Neurochem. 2006 Dec;99(6):1517-30. doi: 10.1111/j.1471-4159.2006.04196.x. Epub 2006 Oct 24.

Abstract

Both cyclic AMP (cAMP) and nerve growth factor (NGF) have been shown to cause rapid activation of cAMP response element-binding protein (CREB) by phosphorylation of serine 133, but additional regulatory events contribute to CREB-targeted gene expression. Here, we have used stable transfection with a simple cAMP response element (CRE)-driven reporter to address the kinetics of CRE-dependent transcription during neuronal differentiation of PC12 cells. In naive cells, dibutyryl cAMP (dbcAMP) generated a rapid increase in CRE-driven luciferase activity by 5 h that returned to naive levels by 24 h. Luciferase induction after NGF treatment was delayed until 48 h when CRE-driven luciferase expression became TrkA dependent. Blocking histone deacetylase (HDAC) activity accelerated NGF-dependent CRE-driven luciferase expression by at least 24 h and resulted in a sustained cAMP-dependent expression of CRE-driven luciferase beyond 24 h. Inhibition of protein synthesis before stimulation with NGF or dbcAMP indicated that both stimuli induce expression of a transcriptional repressor that delays NGF-dependent and attenuates cAMP-dependent CRE-driven transcription. NGF caused a rapid but transient HDAC-dependent increase in inducible cAMP element repressor (ICER) expression, but ICER expression was sustained with increased cAMP. Depletion of ICER from PC12 cells indicated that HDAC-dependent ICER induction is responsible for the delay in CRE-dependent transcription after NGF treatment.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bucladesine / pharmacology
  • Carbazoles / pharmacology
  • Cell Differentiation / drug effects
  • Chromatin Immunoprecipitation / methods
  • Cyclic AMP / pharmacology*
  • Cyclic AMP Response Element Modulator / pharmacology*
  • Cyclic AMP Response Element-Binding Protein / genetics
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Drug Interactions
  • Electrophoretic Mobility Shift Assay / methods
  • Enzyme Inhibitors / pharmacology
  • Gene Expression / drug effects*
  • Gene Expression / physiology
  • Gene Expression Regulation / drug effects*
  • Immunoprecipitation / methods
  • Indole Alkaloids
  • Luciferases / metabolism
  • Nerve Growth Factor / pharmacology*
  • PC12 Cells / drug effects
  • RNA, Messenger / metabolism
  • RNA, Small Interfering / pharmacology
  • Rats
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Time Factors
  • Transfection / methods

Substances

  • Carbazoles
  • Cyclic AMP Response Element-Binding Protein
  • Enzyme Inhibitors
  • Indole Alkaloids
  • RNA, Messenger
  • RNA, Small Interfering
  • Cyclic AMP Response Element Modulator
  • Bucladesine
  • Nerve Growth Factor
  • staurosporine aglycone
  • Cyclic AMP
  • Luciferases