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Vol. 273, Issue 4, C1267-C1277, October 1997
cotransporter
Renal Division, Department of Medicine, Brigham and Women's Hospital, and Harvard Center for the Study of Kidney Diseases, Harvard Medical School, Boston, Massachusetts 02115
The Slc12a2 gene encodes a
widely expressed bumetanide-sensitive
Na+-K+-2Cl
cotransporter that participates in various functions such as Cl secretion and cell
volume regulation. We isolated and characterized 75 kilobases of the
murine gene encoding the cotransporter. The cotransport protein is
encoded by 27 exons. Ribonuclease protection assay and primer extension
demonstrated tissue-specific transcription initiation sites located
within 270 base pairs upstream of the start codon. Nucleotide sequence
analysis of the proximal 5'-flanking region revealed the presence
of a weak TATA box, multiple Sp1/GC consensus sites, and the consensus
sequence of a putative transcriptional initiator. Transfection of
luciferase reporter gene constructs in mouse inner medullary collecting
duct (mIMCD-3) cells confirmed the location of the minimal promoter
within a 120-base pair fragment upstream of the cDNA. We also report
the identification of an alternatively spliced variant of the
cotransporter, expressed primarily in brain. This new spliced variant
lacks exon 21, which encodes a 16-amino acid peptide located in the
COOH-terminal tail of the protein. The absence of this exon causes the
loss of the single protein kinase A consensus site of the cotransport
protein.
promoter; spliced variant; brain
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