Isolation and functional characterization of the 5'-upstream region of mouse P/Q-Type Ca2+ channel α(1A) subunit gene

Eiki Takahashi, Yoshiyuki Murata, Tohru Oki, Norimasa Miyamoto, Yasuo Mori, Naoyuki Takada, Hiroshi Wanifuchi, Nobuko Wanifuchi, Ken ichi Yagami, Tetsuhiro Niidome, Isao Tanaka, Koichi Katayama

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14 Citations (Scopus)

Abstract

The ω-agatoxin-IVA-sensitive P/Q-type Ca2+ channel is predominantly expressed in the nervous system. To dissect the molecular mechanisms underlying the neuron-specific expression of the P/Q-type channel, we have isolated and characterized the 5'-upstream region of the mouse α(1A) subunit gene. A transcription start site appeared to exist at -269 bp upstream from the start codon as found by 5' RACE analysis. The proximal promoter of the α(1A) subunit gene lacks a typical TATA box, but contains several transcription factor binding sequences, including two Sp1 sites. When linked to a placental alkaline phosphatase (FLAP) reporter gene to examine the promoter activity, the 6.3-kb (-6273 to +269) 5'-upstream region, but not a smaller 3.0-kb construct (-3021 to +269), was able to drive the reporter gene in neuron-like PC12 cells. In contrast, neither of these constructs enhanced the FLAP expression in fibroblast NIH3T3 cells. The sequence between 6.3 and 3.0 kb of the 5'-upstream region did not show promoter activity in either of the cell lines, but enhanced TK promoter activity in PC12 cells, though not in NIH3T3 cells. These results suggest that neuron-specific elements of the α(1A) subunit gene are likely to be located in the distal upstream regions(-6273 to -3021) of the 5'-upstream sequence.

Original languageEnglish
Pages (from-to)54-59
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume260
Issue number1
DOIs
Publication statusPublished - Jun 24 1999
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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