Transcriptional regulation of the neuronal L-type calcium channel α1D subunit gene

Timothy J. Kamp, Michael Mitas, Kay L. Fields, Sadamitsu Asoh, Hemin Chin, Eduardo Marban, Marshall Nirenberg

Research output: Contribution to journalArticle

Abstract

1. The transcriptional regulation of the rat brain L-type calcium channel α1D subunit (RBα1D) gene was investigated using NG108-15 neuroblastoma-glioma cells. 2. Differentiation of NG108-15 cells in the presence of prostaglandin E1 or retinoic acid resulted in the appearance of mRNA encoding the RBα1D subunit detected using Northern blot analysis. 3. A rat genomic DNA library was screened, and a 15.2-kb clone was isolated and partially sequenced which included part of the 5′ upstream sequence through the initial part of intron 2 of the RBα1D gene. 4. Deletion analysis, using a CAT reporter gene and transfected NG108-15 cells, revealed that the 1.2-kb 5′-upstream sequence from the RBα1D gene contains cis-acting positive and negative regulatory elements. A deletion of the 3′ end of exon 1 also suggested the presence of regulatory elements in the first exon. 5. DNase footprinting of exon 1 of the RBα1D gene revealed two regions protected from digestion by specific protein binding, and the second region included an (ATG)7 trinucleotide repeat sequence. Electrophoretic mobility shift assays confirmed nuclear protein(s) binding to the (ATG)7 sequence. 6. The (ATG)7 sequence functions as a enhancer when linked to a thymidine kinase promoter and a CAT reporter gene. 7. These results provide the initial description of the transcriptional regulation of the RBα1D gene and identify a novel enhancer that consists of an (ATG)7 trinucleotide repeat sequence.

Original languageEnglish (US)
Pages (from-to)307-326
Number of pages20
JournalCellular and Molecular Neurobiology
Volume15
Issue number3
DOIs
StatePublished - Jun 1995

Keywords

  • calcium channels
  • enhancer element
  • gene expression regulation
  • metal response element
  • molecular cloning
  • NG108-15 cells
  • transcription
  • trinucleotide repeat

ASJC Scopus subject areas

  • Neuroscience(all)
  • Genetics
  • Clinical Biochemistry
  • Cell Biology

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  • Cite this

    Kamp, T. J., Mitas, M., Fields, K. L., Asoh, S., Chin, H., Marban, E., & Nirenberg, M. (1995). Transcriptional regulation of the neuronal L-type calcium channel α1D subunit gene. Cellular and Molecular Neurobiology, 15(3), 307-326. https://doi.org/10.1007/BF02089942