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Am J Physiol Renal Physiol 283: F904-F911, 2002. First published July 9, 2002; doi:10.1152/ajprenal.00156.2002
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Vol. 283, Issue 5, F904-F911, November 2002

NF-kappa B inhibits transcription of the H+-K+-ATPase alpha 2-subunit gene: role of histone deacetylases

Wenzheng Zhang and Bruce C. Kone

Departments of Internal Medicine and of Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, Houston, Texas 77030

The H+-K+-ATPase alpha 2 (HKalpha 2) gene plays a central role in potassium homeostasis, yet little is known about its transcriptional control. We recently demonstrated that the proximal promoter confers basal transcriptional activity in mouse inner medullary collecting duct 3 cells. We sought to determine whether the kappa B DNA binding element at -104 to -94 influences basal HKalpha 2 gene transcription in these cells. Recombinant NF-kappa B p50 footprinted the region -116/-94 in vitro. Gel shift and supershift analysis revealed NF-kappa B p50- and p65-containing DNA-protein complexes in nuclear extracts of mouse inner medullary collecting duct 3 cells. A promoter-luciferase construct with a mutated -104/-94 NF-kappa B element exhibited higher activity than the wild-type promoter in transfection assays. Overexpression of NF-kappa B p50, p65, or their combination trans-repressed the HKalpha 2 promoter. The histone deacetylase (HDAC) inhibitor trichostatin A partially reversed NF-kappa B-mediated trans-repression of the HKalpha 2 promoter. HDAC6 overexpression inhibited HKalpha 2 promoter activity, and HDAC6 coimmunoprecipitated with NF-kappa B p50 and p65. These results suggest that HDAC6, recruited to the DNA protein complex, acts with NF-kappa B to suppress HKalpha 2 transcription and identify NF-kappa B p50 and p65 as novel binding partners for HDAC6.

kidney; colon; trichostatin A; promoter


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