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Cloning and characterization of promoter and 5'-UTR of the NMDA receptor subunit ε2: evidence for alternative splicing of 5'-non-coding exon

Klein, M., Pieri, I., Uhlmann, F., Pfizenmaier, K. & Eisel, U., 1998, In : Gene. 208, 2, p. 259-269 11 p.

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  • Matthias Klein
  • Isabelle Pieri
  • Frank Uhlmann
  • Klaus Pfizenmaier
  • Ulrich Eisel
Using rapid amplification of cDNA ends (RACE), we have cloned the 5'-untranslated region (5'-UTR) of the N-methyl-D-aspartate receptor subunit ε2 from murine forebrain-derived mRNA. We identified two distinct types of cDNA species differing in the presence or absence of one exon sequence. Sequencing of the 5'-non-coding region of the ε2 gene revealed that the ε2 5'-UTR consists of three untranslated exons located at least 20 kb upstream of exon 4 that contains the ATG codon for initiation of translation. This genomic organization shows a close similarity to the ε3 gene. The transcriptional start site was determined by primer extension assays. Expression of the alternative exon sequence was shown by in situ hybridization in the murine brain. Basal transcriptional activity of the ε2 promoter was detected in different neuronal and non-neuronal cell lines with transient reporter gene expression assays. Potential SP1 and CREB binding sites were found in the promoter region. Specific binding of these transcription factors was demonstrated in electrophoretic mobility shift assays.
Original languageEnglish
Pages (from-to)259-269
Number of pages11
JournalGene
Volume208
Issue number2
Publication statusPublished - 1998

    Keywords

  • Rapid amplification of cDNA ends, Sp1, CREB, Upstream open reading frame, Glutamate receptors

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