Abstract
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 language | English |
|---|---|
| Pages (from-to) | 2067-2075 |
| Number of pages | 9 |
| Journal | Journal of Neurochemistry |
| Volume | 59 |
| Issue number | 6 |
| Publication status | Published - Dec 1992 |
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Keywords
- c-Fos
- Cerebellar granule cells
- DNA-binding activity
- Glutamate receptors
- Kainate
- N-Methyl-D-aspartate
ASJC Scopus subject areas
- Biochemistry
- Cellular and Molecular Neuroscience
Cite this
Stimulation of cultured cerebellar granule cells via glutamate receptors induces TRE- and CRE-binding activities mediated by common DNA-binding complexes. / Sakurai, Hiroaki; Kurusu, Rie; Sano, Kuniaki; Tsuchiya, Tomofusa; Tsuda, Masaaki.
In: Journal of Neurochemistry, Vol. 59, No. 6, 12.1992, p. 2067-2075.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Stimulation of cultured cerebellar granule cells via glutamate receptors induces TRE- and CRE-binding activities mediated by common DNA-binding complexes
AU - Sakurai, Hiroaki
AU - Kurusu, Rie
AU - Sano, Kuniaki
AU - Tsuchiya, Tomofusa
AU - Tsuda, Masaaki
PY - 1992/12
Y1 - 1992/12
N2 - 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.
AB - 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.
KW - c-Fos
KW - Cerebellar granule cells
KW - DNA-binding activity
KW - Glutamate receptors
KW - Kainate
KW - N-Methyl-D-aspartate
UR - http://www.scopus.com/inward/record.url?scp=0026498429&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0026498429&partnerID=8YFLogxK
M3 - Article
VL - 59
SP - 2067
EP - 2075
JO - Journal of Neurochemistry
JF - Journal of Neurochemistry
SN - 0022-3042
IS - 6
ER -