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1 McGill University-Montreal Children's Hospital Research Institute and Departments of 2 Pediatrics and 3 Human Genetics, McGill University, Montreal, Quebec, Canada H3Z 2Z3
First published August 15, 2001;
10.1152/ajprenal.00092.2001.
Na-phosphate (Pi)
cotransporters in the apical membrane of renal proximal tubular cells
play a major role in the maintenance of Pi homeostasis.
Although two such cotransporters, Npt1 and Npt2, have been
identified, little is known about the function and regulation of Npt1.
We cloned and characterized the murine (Npt1) and human
(NPT1) genes, isolated the 5'-flanking region of
Npt1, and analyzed its promoter activity. Npt1 is
~29 kb with 12 exons, whereas NPT1 is ~49 kb with one
additional exon. The Npt1 promoter has a TATA-like box but
no CAAT box, and the transcription start site was identified by primer
extension and 5'-rapid amplification of cDNA ends. Transfection of
opossum kidney cells with Npt1 promoter-reporter gene
constructs demonstrated significant activity in a 570-bp fragment that
was completely inhibited by cotransfection with the transcription
factor, hepatocyte nuclear factor (HNF)-3
. Deletion of 200 bp from
the 3'-end of the 570-bp fragment abrogated its promoter activity. In
addition, promoter activity of a 4.5-kb fragment, but not the 570-bp
fragment, was stimulated fourfold by cotransfection with HNF-1
.
Other well-characterized cis-acting elements were identified in the
Npt1 promoter. We suggest that Npt1 expression is
transcriptionally regulated and provide a basis for the investigation
of Npt1 function by targeted mutagenesis.
kidney; brush-border membrane; hepatocyte nuclear factor; transcription
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