Characterization of the rabbit HKalpha2 gene promoter.

The HKalpha2 gene directs synthesis of the HKalpha2 subunit of the H(+), K(+)-ATPase. In the kidney and colon, the gene is highly expressed and is thought to play a role in potassium (K(+)) conservation. The rabbit has been an important experimental system for physiological studies of ion transport in the kidney, so the rabbit HKalpha2 gene has been cloned and characterized. The genomic clones and the previously reported HKalpha2a and HKalpha2c subunit cDNAs provided a means to address several issues regarding the structure and expression of the HKalpha2 gene. First, the genomic organization established that the rabbit HKalpha2 gene was unambiguously homologous to the mouse HKalpha2 gene and the human ATP1AL1 gene. Second, the mapping of the transcription start site for the alternate transcript, HKalpha2c, confirmed that it was an authentic rabbit transcript. Finally, isolation of DNA from the 5' end of the HKalpha2 gene enabled us to initiate studies on its regulation in the rabbit cortical collecting duct. The promoter and two putative negative regulatory regions were identified and the effect of cell confluency on gene expression was studied.