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Volume 272, Number 9, Issue of February 28, 1997 pp. 5936-5942
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Nerve Growth Factor Up-regulates the N-Methyl-D-aspartate Receptor Subunit 1 Promoter in PC12 Cells

(Received for publication, July 29, 1996, and in revised form, November 1, 1996)

Guang Bai and John W. Kusiak

From the Molecular Neurobiology Unit, Gerontology Research Center, NIA, National Institutes of Health, Baltimore, Maryland 21224

The N-methyl-D-aspartate (NMDA) subtype of glutamate receptor plays important roles in synaptic plasticity, the induction of long term potentiation, and excitotoxicity. Mechanisms governing the regulation of expression of its subunit genes remain largely unknown. The promoter of the essential subunit of the NMDA receptor heteromer, NMDAR1, contains DNA binding elements recognized by the nerve growth factor-inducible/early growth reaction factor (NGFI/Egr) family of transcription factors that are rapidly induced by neurotrophins, such as nerve growth factor (NGF). This study examined the effect of NGF on the activity of the N-methyl-D-aspartate receptor subunit 1 (NMDAR1) promoter/luciferase reporter constructs in PC12 cells, which contain the high affinity TrkA receptor for NGF and the low affinity p75NTR receptor for neurotrophins. NGF up-regulated the activity of the NMDAR1 promoter by 3-4-fold in a time- and dose-dependent manner. 5' deletional analysis of the promoter indicated that the responsive element(s) resides in the proximal region containing GSG and Sp1 sites. Mutational analysis of these sites revealed that both were important for NGF regulation. Transient expression of Egr-1 increased activity of the wild type promoter but failed to increase activity of a GSG mutant promoter. Other neurotrophins did not activate the promoter, while K-252a inhibited the action of NGF. These results suggest that the NGF effect is mediated by the high affinity NGF receptor, Trk A and that neurotrophin binding to the low affinity neurotrophin receptor, p75NTR, alone does not affect the promoter activity. Our results suggest that NGF is able to up-regulate the activity of the NMDAR1 promoter and may play a role in controlling the expression levels of NMDA receptors.


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