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J. Biol. Chem., Vol. 276, Issue 28, 26260-26268, July 13, 2001
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From the Division of Nephrology, Endocrinology, and Vascular
Medicine, Department of Medicine, Tohoku University Graduate School of
Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan
The molecular mechanism underlying the renal
expression localization of the thiazide-sensitive Na-Cl
cotransporter (TSC) gene was studied. The
TSC gene was localized to chromosome 19p12-14. In
cultured cells, tissue-specific transcription activity of the 5'-flanking region of the rat rTSC gene (5'FL/rTSC) was
demonstrated, and the major promoter region was located between
position The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AB024534.
Renal Tubule-specific Transcription and Chromosomal Localization
of Rat Thiazide-sensitive Na-Cl Cotransporter Gene*
,,
580 and 141. To further examine the tissue-specific
transcription, transgenic rats harboring the 5'FL/rTSC fused upstream
of the LacZ gene were generated. Immunohistochemical
analysis clearly showed that LacZ gene expression was
co-localized to distal convoluted tubules (DCT) with TSC, indicating
that the 5'FL/rTSC regulates the renal tubule-specific TSC expression.
Because a transcription factor, HFH-3 (hepatocyte nuclear factor-3/folk
head homologue-3), had also been localized to DCT, a possible role of
the putative cis-acting element (HFH-3/rTSC, 400/387
position) for HFH-3 binding in the tissue-specific transcription was
examined. Deletion and mutation analyses suggested that transcription
of the HFH-3/rTSC was actually responsive to HFH-3, and electrophoretic
mobility shift assay showed a direct binding of in vitro
synthesized HFH-3 to the HFH-3/rTSC. In conclusion, the
rTSC gene is localized to rat chromosome 19p12-24. The
transcription regulatory region of the TSC gene confers
DCT-specific gene expression. DCT-specific transcription factor HFH-3
may be involved in the renal tubule-specific transcription of
TSC gene.
*
This study was supported in part by grants-in-aid from the
Ministry of Education, Science, and Culture, Japan and by the Takeda Foundation for Metabolic Disorder Research, Japan.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
These authors were equal contributors to this study.
§
To whom correspondence should be addressed: Molecular Biology Unit,
Division of Nephrology, Endocrinology, and Vascular Medicine, Dept. of
Medicine, Tohoku University Graduate School of Medicine, 1-1 Seiryo-machi, Aoba-ku, Sendai 980-8574, Japan. Tel.: 81-22-717-7163; Fax: 81-22-717-7168; E-mail: kazut2i@mail.cc.tohoku.ac.jp.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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