HNF-1B specifically regulates the transcription of the γa-subunit of the Na+/K+-ATPase

► Defects in HNF-1B transcription factor affect Mg2+ handling in the distal kidney. ► γa- and γb- subunits of the Na+/K+-ATPase colocalize in the distal convoluted tubule of the nephron. ► HNF-1B specifically activates γa expression. ► HNF-1B mutants have a dominant negative effect on wild type HNF-1B activity. ► Defective transcription of γa may promote renal Mg2+ wasting. Hepatocyte nuclear factor-1B (HNF-1B) is a transcription factor involved in embryonic development and tissue-specific gene expression in several organs, including the kidney. Recently heterozygous mutations in the HNF1B gene have been identified in patients with hypomagnesemia due to renal Mg2+ wasting. Interestingly, ChIP–chip data revealed HNF-1B binding sites in the FXYD2 gene, encoding the γ-subunit of the Na+/K+-ATPase. The γ-subunit has been described as one of the molecular players in the renal Mg2+ reabsorption in the distal convoluted tubule (DCT). Of note, the FXYD2 gene can be alternatively transcribed into two main variants, namely γa and γb. In the present study, we demonstrated via two different reporter gene assays that HNF-1B specifically acts as an activator of the γa-subunit, whereas the γb-subunit expression was not affected. Moreover, the HNF-1B mutations H69fsdelAC, H324S325fsdelCA, Y352finsA and K156E, previously identified in patients with hypomagnesemia, prevented transcription activation of γa-subunit via a dominant negative effect on wild type HNF1-B. By immunohistochemistry, it was shown that the γa- and γb-subunits colocalize at the basolateral membrane of the DCT segment of mouse kidney. On the basis of these data, we suggest that abnormalities involving the HNF-1B gene may impair the relative abundance of γa and γb, thus affecting the transcellular Mg2+ reabsorption in the DCT.