Isolation and characterization of kidney-specific ClC-K1 chloride channel gene promoter

The American Journal of Physiology
S UchidaF Marumo

Abstract

The rat ClC-K1 chloride channel is a kidney-specific member of the ClC chloride channel family found exclusively in the thin ascending limb of Henle's loop in the kidney. To gain insight into the mechanism(s) of kidney-specific expression of ClC-K1, a genomic clone that contains the 5'-flanking region of the rat ClC-K1 gene was isolated. A single transcription start site was located 84 bp upstream of the start codon. The sequence of the proximal 5'-flanking region contained an activator protein (AP)-3 site, a glucocorticoid-responsive element, several AP-2 sites, and several E-boxes, but it lacked a TATA box. To functionally express the promoter, the approximately 2.5-kb pair 5'-flanking region was ligated to a luciferase reporter gene and transfected into inner medullary (IM) cells, a stable ClC-K1-expressing cell line derived from the inner medulla of simian virus 40 transgenic mouse, and ClC-K1-nonexpressing cell lines. Luciferase activity was 7-to 24-fold greater in IM cells than those in nonexpressing cell lines, suggesting that the approximately 2.5-kb fragment contained cis-acting regulatory elements for cell-specific expression of the ClC-K1 gene. Deletion analysis revealed that this cell-specific promoter activity in I...Continue Reading

References

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Feb 14, 1995·Proceedings of the National Academy of Sciences of the United States of America·M TakenakaJ S Handler
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Citations

Mar 7, 2008·American Journal of Physiology. Cell Physiology·Lior AdlerIsrael Zelikovic
Dec 21, 2007·Nature Clinical Practice. Nephrology·Bernhard K KrämerSiegfried Waldegger
Mar 7, 2002·American Journal of Physiology. Renal Physiology·Xinhua ZhuXue-Ru Wu
Mar 28, 2002·Physiological Reviews·Thomas J JentschAnselm A Zdebik
Sep 9, 2004·American Journal of Physiology. Renal Physiology·Edna EfratiIsrael Zelikovic
May 31, 2018·Physiological Reviews·Thomas J Jentsch, Michael Pusch

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