Intracellular zinc-dependent TAS2R8 gene expression through CTCF activation.

Tsuyoshi Kojima, Toyonobu Maeda, Atsuko Suzuki, Tetsuo Yamamori, Yasumasa Kato

Biomedical research (Tokyo, Japan) 2020

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Taste-2 receptors (TAS2Rs), which belong to the G-protein coupled receptor (GPCR) family, are receptors for bitter taste perception. The aim of this study was to investigate whether zinc deficiency affects the expression of TAS2R genes. The promoter activity of the TAS2R7, TAS2R8, and TAS2R42 genes were determined in Ca9-22 oral squamous cell carcinoma cells cultured in the presence or absence of zinc. Luciferase reporter assays showed that zinc deprivation inhibited TAS2R8 promoter activity, but not the promoter activity of the other two genes. Treatment of the cells with N,N,N',N'-tetrakis(2-pyridinylmethyl)-1,2-ethanediamine (TPEN), an intracellular chelator of Zn2+, in the presence of 10% fetal bovine serum reduced TAS2R8 promoter activity. Truncation/deletion mutants of TAS2R8 promoter-luciferase constructs showed that the region from nucleotide -1152 to nucleotide -925 was critical for intracellular zinc dependency and contained a CCCTC-binding factor (CTCF) binding motif. A chromatin immunoprecipitation (ChiP) assay showed that CTCF bound specifically to this region, a binding abrogated by zinc deficiency, suggesting that CTCF plays a critical role in zinc-dependent bitter taste perception through TAS2R8.

Citation

Tsuyoshi Kojima, Toyonobu Maeda, Atsuko Suzuki, Tetsuo Yamamori, Yasumasa Kato. Intracellular zinc-dependent TAS2R8 gene expression through CTCF activation. Biomedical research (Tokyo, Japan). 2020;41(5):217-225