Stimulation of cultured cerebellar granule cells via glutamate receptors induces TRE- and CRE-binding activities mediated by common DNA-binding complexes

Hiroaki Sakurai, Rie Kurusu, Kuniaki Sano, Tomofusa Tsuchiya, Masaaki Tsuda

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By use of nuclear mini-extracts prepared from cultured cerebellar granule cells in a gel-mobility assay, exogenous N-methyl-D-aspartate (NMDA) or kainate was shown to increase both 12-O-tetradecanoylphorbol 13-acetate-responsive element (TRE)- and cyclic AMP-responsive element (CRE)-binding activity. These increases were specifically prevented by the NMDA receptor antagonist D,L-2-amino-5-phosphonovalerate and the non-NMDA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione, respectively. The increase of TRE-binding activity was dependent on de novo protein synthesis, and its inductions by both NMDA and kainate required extracellular Ca2+. TRE-binding activity was competitively inhibited by the CRE, and vice versa, showing higher DNA-binding affinity to the CRE than to the TRE. A proteolytic clipping bandshift assay demonstrated that the increase in CRE-binding activity could be mediated by the TRE-binding activity. Thus, the TRE-binding activity cross-binding to the CRE could be activated by NMDA or kainate stimulation. The involvement of c-Fos or Fos-related proteins in the TRE- and CRE-binding complexes was shown by a supershift gel-mobility assay using anti-c-Fos antiserum.

Original languageEnglish
Pages (from-to)2067-2075
Number of pages9
JournalJournal of Neurochemistry
Issue number6
Publication statusPublished - Dec 1992



  • c-Fos
  • Cerebellar granule cells
  • DNA-binding activity
  • Glutamate receptors
  • Kainate
  • N-Methyl-D-aspartate

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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