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J. Biol. Chem., Vol. 277, Issue 47, 44911-44919, November 22, 2002

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Neuronal B-binding Factors Consist of Sp1-related Proteins
FUNCTIONAL IMPLICATIONS FOR AUTOREGULATION OF N-METHYL-D-ASPARTATE RECEPTOR-1 EXPRESSION^*

Xianrong Mao, Andrea M. Moerman^§, and Steven W. Barger^§¶

From the Departments of  Anatomy and Neurobiology and ^§ Geriatrics, University of Arkansas for Medical Sciences and the ^¶ Geriatric Research Education and Clinical Center, Central Arkansas Veterans Healthcare System, Little Rock, Arkansas 72205

Neurons contain a protein factor capable of binding DNA elements normally bound by the transcription factor NF-B. However, several lines of evidence suggest that this neuronal B-binding factor (NKBF) is not bona fide NF-B. We have identified NKBF from cultures of neocortical neurons as a complex containing proteins related to Sp1. This complex was bound by antibodies to Sp1, Sp3, and Sp4 and was competed from binding to an NF-B element by an oligonucleotide containing an Sp1-binding site. This Sp1 oligonucleotide detected an abundant factor in neuronal nuclei that migrated in electrophoretic mobility shift assays at a position consistent with NKBF. Expression of transfected Sp1 stimulated transcription in a manner dependent upon a B cis-element. Similar to our previous reports for NKBF (Mao, X., Moerman, A. M., Lucas, M. M., and Barger, S. W. (1999) J. Neurochem. 73, 1851-1858 and Moerman, A. M., Mao, X., Lucas, M. M., and Barger, S. W. (1999) Mol. Brain Res. 67, 303-315), the activity of the Sp1-related factor was reduced by activation of ionotropic glutamate receptors, consistent with proteolytic degradation of all three Sp1-related factors. Expression of the N-methyl-D-aspartate receptor-1 (NR1) subunit of glutamate receptors correlated with the activity of the Sp1-related factor, specifically through an Sp1 element in the NR1 promoter. These data provide the first evidence that Sp1 or related family members are responsible for B-binding activity and are involved in a negative feedback for NR1 in central nervous system neurons.

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