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Regulation of the murine NMDA‐receptor‐subunit NR2C promoter by Sp1 and fushi tarazu factor1 (FTZ‐F1) homologues
Author(s) -
Pieri Isabelle,
Klein Matthias,
Bayertz Christian,
Gerspach Jeannette,
van der Ploeg Anja,
Pfizenmaier Klaus,
Eisel Ulrich
Publication year - 1999
Publication title -
european journal of neuroscience
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.346
H-Index - 206
eISSN - 1460-9568
pISSN - 0953-816X
DOI - 10.1046/j.1460-9568.1999.00629.x
Subject(s) - microbiology and biotechnology , biology , promoter , reporter gene , transcription factor , electrophoretic mobility shift assay , transcription (linguistics) , binding site , point mutation , gene , gene expression , sp1 transcription factor , transcriptional regulation , mutant , genetics , linguistics , philosophy
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.