Cloning of the Dopamine-1A(D_<1A>) Receptor Gene Expressed in Porcine Renal Epithelial Cells

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Author(s)

    • HEALY Dennis P.
    • Department of Pharmacology, Mount Sinai School of Medicine of the City University of New York
    • O'ROURKE Dawn A.
    • Department of Pharmacology, Mount Sinai School of Medicine of the City University of New York
    • GRENADER Arcady C.
    • Department of Pharmacology, Mount Sinai School of Medicine of the City University of New York

Abstract

We sought to determine the molecular identify of the dopamine-1 (D<sub>1</sub>) receptor expressed in the porcine renal epithelial cell line LLC-PK<sub>1</sub>. We first isolated a partial cDNA by the reverse transcription- polymerase chain reaction procedure and then used the partial cDNA to isolate positive overlapping clones from a porcine genomic DNA library. Sequence analysis of the gene revealed that the longest open-reading frame encoded a 446 amino acid protein that was 95% identical to the human D<sub>1A</sub> receptor. Expression studies in mammalian cells were also consistent with the clones encoding a D<sub>1</sub> receptor. Northern blot hybridizations with LLC-PK<sub>1</sub> poly (A<sup>+</sup>) RNA were strongly positive. The porcine D<sub>1A</sub> gene has two exons and a short intron in the 5′ untranslated region. The 5′ flanking region lacks a TATA and CAAT box but is high in GC content (68%) and contains multiple Sp1 binding sites. The 5′ flanking region also contains numerous other <i>cis</i>-acting elements for transcription factors. These results indicate that the D<sub>1A</sub> receptor is the major D<sub>1</sub> receptor expressed in LLC-PK<sub>1</sub> cells and further suggest that LLC-PK<sub>1</sub> cells may be a useful model to study the regulation of renal D<sub>1A</sub> receptor gene transcription. (<i>Hypertens Res</i> 1995; 18 Suppl. I: S11-S17)

Journal

  • Hypertension Research

    Hypertension Research 18, S11-S17, 1995-06-01

    The Japanese Society of Hypertension

References:  28

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