Regulation of the murine NMDA-receptor-subunit NR2C promoter by Sp1 and fushi tarazu factor1 (FTZ-F1) homologues

We have cloned the 5′‐region of the murine N‐methyl‐d‐aspartate (NMDA) receptor channel subunit NR2C (GluRε3) gene and characterized the cis‐ and trans‐activating regulatory elements responsible for its tissue specific activity. By using a native ε3‐promoter/lacZ‐construct & various 5′‐deletion...

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Bibliographic Details
Published in:	The European journal of neuroscience Vol. 11; no. 6; pp. 2083 - 2092
Main Authors:	Pieri, Isabelle, Klein, Matthias, Bayertz, Christian, Gerspach, Jeannette, van der Ploeg, Anja, Pfizenmaier, Klaus, Eisel, Ulrich
Format:	Journal Article
Language:	English
Published:	Oxford, UK Blackwell Science Ltd 01-06-1999 Wiley Subscription Services, Inc
Subjects:	3T3 Cells 5′-untranslated region Animals Base Sequence - genetics Cell Line cell type specific expression Cloning Culture Techniques DNA-Binding Proteins - physiology Fushi Tarazu Transcription Factors Gene expression Gene Expression - physiology gene regulation Homeodomain Proteins Isomerism Lac Operon - genetics Mice Molecular Sequence Data Mutation NMDAR2C (ε3) orphan nuclear hormone receptor Plasmids Polymerase chain reaction Promoter Regions, Genetic - genetics Promoter Regions, Genetic - physiology Receptors, Cytoplasmic and Nuclear Receptors, N-Methyl-D-Aspartate - genetics Receptors, N-Methyl-D-Aspartate - metabolism Sp1 Transcription Factor - physiology Steroidogenic Factor 1 transcription factors Transcription Factors - metabolism Transcription Factors - physiology Germany
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Summary:	We have cloned the 5′‐region of the murine N‐methyl‐d‐aspartate (NMDA) receptor channel subunit NR2C (GluRε3) gene and characterized the cis‐ and trans‐activating regulatory elements responsible for its tissue specific activity. By using a native ε3‐promoter/lacZ‐construct & various 5′‐deletion constructs, we compared β‐galactosidase expression in non‐neuronal NIH3T3 cells and in neuronal ε3‐gene‐expressing HT‐4 cells and show that large parts of the ε3 promoter are responsible for the repression of the ε3 gene in non‐neuronal cells. Deletion of exon 1 sequences led to an enhancement of ε3 transcription, suggesting a role of the 5′‐untranslated region in ε3 gene regulation. Sequence analysis of the promoter region revealed potential binding sites for the transcription factor Sp1, the murine fushi tarazu factor1 (FTZ‐F1) homologues, embryonic LTR binding proteins (ELP1,2,3) and steroidogenic factor (SF‐1), as well as for the chicken ovalbumin upstream promoter transcription‐factor (COUP‐TF). Electrophoretic mobility shift assays confirmed specific binding of Sp1, SF‐1 and COUP‐TFI. Whereas point mutation studies indicate that, in neuronal HT‐4 cells, Sp1 is apparently not critically involved in basal ε3 gene transcription, SF1 is a positive regulator. This was evident from a selective enhancement of ε3‐promoter‐driven reporter gene expression upon cotransfection of an SF1‐expression vector, which was reverted by deletion and point mutation of the SF1 binding site.
Bibliography:	istex:A426A874D6DA51ADA44C0C1C87F7ACF174309BA1 ark:/67375/WNG-6975NZ04-7 ArticleID:EJN629 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0953-816X 1460-9568
DOI:	10.1046/j.1460-9568.1999.00629.x