Identification of a rapidly dephosphorylating 95-kDa protein as elongation factor 2 during 8-Br-cAMP treatment of N1E115 neuroblastoma cells

Biochem Biophys Res Commun. 1995 Dec 5;217(1):131-7. doi: 10.1006/bbrc.1995.2754.

Abstract

Treatment of 8-Br-cAMP promotes neurite outgrowth and neuronal differentiation in N1E115 mouse neuroblastoma cells. Prior or simultaneous treatment of PMA blocks 8-Br-cAMP-mediated neurite outgrowth. Phosphorylation of cellular proteins during these treatments was examined in a permeabilized cell system. While PMA promotes phosphorylation of the heat-stable protein kinase C substrates MARCKS and neuromodulin, 8-Br-cAMP hastens the dephosphorylation of a protein of M(r)95k (p95). Extensively purified, N-terminal sequenced, and judged from its phosphorylation properties, p95 was identified as the eukaryotic elongation factor-2 (eEF-2), whose dephosphorylation has been reported to be related to an increase in protein synthesis. It is likely 8-Br-cAMP stimulates dephosphorylation of eEf-2, promotes protein synthesis that eventually leads to neuronal differentiation in N1E115 cells.

MeSH terms

  • 8-Bromo Cyclic Adenosine Monophosphate / pharmacology
  • Amino Acid Sequence
  • Animals
  • Cell Differentiation / drug effects
  • Cyclic AMP-Dependent Protein Kinases / metabolism
  • Mice
  • Molecular Sequence Data
  • Molecular Weight
  • Nerve Tissue Proteins / chemistry
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Neuroblastoma / metabolism*
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / metabolism
  • Peptide Elongation Factor 2
  • Peptide Elongation Factors / chemistry
  • Peptide Elongation Factors / genetics
  • Peptide Elongation Factors / metabolism*
  • Phosphoproteins / chemistry
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Phosphorylation
  • Protein Kinase C / metabolism
  • Tetradecanoylphorbol Acetate / pharmacology
  • Tumor Cells, Cultured

Substances

  • Nerve Tissue Proteins
  • Peptide Elongation Factor 2
  • Peptide Elongation Factors
  • Phosphoproteins
  • 8-Bromo Cyclic Adenosine Monophosphate
  • Cyclic AMP-Dependent Protein Kinases
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate