Demonstration of an E-box and its CNS-related binding factors for transcriptional regulation of the mouse type inositol 1,4,5-trisphosphate receptor gene

Yoshiyuki Konishi, Yasushi Kobayashi, Toshihiko Kishimoto, Yasutaka Makino, Atsushi Miyawaki, Teiichi Furuichi, Hideyuki Okano, Katsuhiko Mikoshiba, Taka Aki Tamura

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The type 1 inositol 1,4,5-trisphosphate receptor (IP3R1) is expressed abundantly in the CNS, such as in cerebellar Purkinje cells and the hippocampus. We established a tissue-specific cell-free transcription system and studied regulatory properties of the 5' upstream region of the IP3R1 gene by use of this system. Deletion analyses of the promoter revealed several cis elements that function significantly in brain nuclear extracts. Among those elements, sequences from -396 to -295 showed the most predominant cerebellum-specific positive function. Footprint analyses demonstrated a factor-binding region from -334 to -318, termed box-I, that contained an E- box consensus sequence. Electrophoretic mobility shift assay revealed CNS- related basic helix-loop-helix proteins for the box-I. Mutational studies using the function assay end competitive electrophoretic mobility shift assays demonstrated a good correlation between the box-I-binding factors and the activated transcription. Box-I-binding factors were present abundantly in adult mouse CNS, whereas their existence was restricted in embryonic and nonneural tissues. Transient chloramphenicol acetyltransferase assay for the IP3R1 promoter revealed the requirement of box-I in Neuro2a neuroblastoma cells. In the postnatal CNS, multiple basic helix-loop-helix factors are expressed abundantly, some of which are suggested to activate IP3R1 gene expression in the mammalian CNS.

Original languageEnglish
Pages (from-to)476-484
Number of pages9
JournalJournal of Neurochemistry
Volume69
Issue number2
Publication statusPublished - 1997 Aug 1
Externally publishedYes

Keywords

  • Basic helix-loop-helix factor
  • E-box
  • In vitro transcription
  • Neuronal gene expression
  • Promoter
  • Type 1 inositol 1,4,5-trisphosphate receptor

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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    Konishi, Y., Kobayashi, Y., Kishimoto, T., Makino, Y., Miyawaki, A., Furuichi, T., Okano, H., Mikoshiba, K., & Tamura, T. A. (1997). Demonstration of an E-box and its CNS-related binding factors for transcriptional regulation of the mouse type inositol 1,4,5-trisphosphate receptor gene. Journal of Neurochemistry, 69(2), 476-484.