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J Am Soc Nephrol 12:2554-2564, 2001
© 2001 American Society of Nephrology

Structure, Promoter Analysis, and Chromosomal Localization of the Murine H+/K+-ATPase {alpha}2 Subunit Gene

Wenzheng Zhang*, Teresa Kuncewicz*, Sandra C. Higham* and Bruce C. Kone*{dagger}

Departments of *Internal Medicine and {dagger}Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, Houston, Texas.

Correspondence to Dr. Bruce C. Kone, Departments of Internal Medicine and Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, 6431 Fannin, MSB 4.138, Houston, TX 77030. Phone: 713-500-6870; Fax: 713-500-6890 (or 6882); E-mail: Bruce.C.Kone{at}uth.tmc.edu

ABSTRACT. The H+/K+-ATPase {alpha}2 subunit (HK{alpha}2) of distal colon and renal collecting ducts plays a critical role in potassium and acid-base homeostasis. The isolation and complete sequence of the murine HK{alpha}2 gene are reported. The HK{alpha}2 gene contains 23 exons and spans 23.5 kb of genomic DNA. The exon/intron organization is comparable to that of the human ATP1AL1 gene. Primer extension and 5'-rapid amplification of cDNA ends of distal colon RNA were used to map the transcription initiation site. Fluorescence in situ hybridization analysis localized the HK{alpha}2 gene to murine chromosome 14C3. Sequence analysis of 7.2 kb of the 5'-flanking region revealed numerous consensus sites for transcription factors, including two potential glucocorticoid response elements. Transient transfection of promoter-luciferase constructs demonstrated strong basal HK{alpha}2 promoter activity in renal collecting duct cells but not in fibroblasts or in a medullary thick ascending limb of Henle’s loop cell line. Deletion analysis revealed that the proximal 0.2 kb of the promoter was sufficient to confer activity in collecting duct cells. These data should prove important in elucidation of the mechanisms controlling the differential, tissue-specific expression of the HK{alpha}2 gene.




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