Phosphorylation-dependent degradation of transgenic CREB protein initiated by heterodimerization

Brain Res. 2007 Jan 26;1130(1):31-7. doi: 10.1016/j.brainres.2006.10.076. Epub 2006 Dec 13.

Abstract

The transcription factor CREB (cyclic AMP response element binding protein) is implicated in diverse brain functions and represents a prospective target in gene therapy for human disorders. However, the transgenic expression and stability of exogenously expressed CREB within the cell remains poorly characterized. Here we found that transient expression of a CREB dominant interfering mutant A-CREB or the inducible cAMP early repressor, ICER, led to the dramatic decrease of exogenously co-expressed CREB in 293 human embryonic kidney cells. Elevation of protein kinase A activity within the cells restored CREB protein levels. A-CREB did not effect the transient expression of a truncated CREB lacking the leucine zipper domain demonstrating a specific effect of heterodimerization on CREB protein stability. Somatic gene transfer into the rat brain using a recombinant adeno-associated virus vector provided robust expression of both transgenic CREB and ICER mRNAs under the control of a constitutive neuron specific enolase (NSE) promoter. In contrast to ICER, the expression of the transgenic CREB mRNA did not result in elevation of CREB protein levels within dentate granule cells of the hippocampus, suggesting its prompt degradation under basal conditions. However, following tetanization of the perforant pathway, which is known to induce CREB phosphorylation, there was a significant increase in the amount of transgenic CREB protein within dentate granule cells. Hence, heterodimerization of unphosphorylated CREB with either A-CREB or ICER triggers CREB protein degradation, whereas phosphorylation prevents CREB from such degradation both in vitro and in vivo.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Genetically Modified
  • Cell Line
  • Cyclic AMP Response Element Modulator / genetics
  • Cyclic AMP Response Element Modulator / metabolism*
  • Cyclic AMP Response Element-Binding Protein / genetics
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Dimerization
  • Electric Stimulation
  • Gene Expression Regulation
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Humans
  • Kidney / cytology
  • Male
  • Neurons / metabolism
  • Perforant Pathway / cytology
  • Perforant Pathway / metabolism*
  • Phosphopyruvate Hydratase / genetics
  • Phosphopyruvate Hydratase / metabolism
  • Phosphorylation
  • Promoter Regions, Genetic
  • RNA, Messenger / analysis
  • Rats
  • Rats, Wistar
  • Recombinant Fusion Proteins / metabolism

Substances

  • A-CREB protein
  • CREM protein, human
  • Cyclic AMP Response Element-Binding Protein
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Cyclic AMP Response Element Modulator
  • Phosphopyruvate Hydratase