Structure, promoter analysis, and chromosomal localization of the murine H(+)/K(+)-ATPase alpha 2 subunit gene.
The H(+)/K(+)-ATPase alpha2 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.
Zhang
Wenzheng
W
Departments of *Internal Medicine and Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, Houston, Texas.
Kuncewicz
Teresa
T
Departments of *Internal Medicine and Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, Houston, Texas.
Higham
Sandra C
SC
Departments of *Internal Medicine and Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, Houston, Texas.
Kone
Bruce C
BC
Departments of *Internal Medicine and Integrative Biology, Pharmacology, and Physiology, The University of Texas Medical School at Houston, Houston, Texas.
D000818:Animals; D001483:Base Sequence / Q000235:genetics; D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D017353:Gene Deletion; D016678:Genome; D017506:H(+)-K(+)-Exchanging ATPase / Q000235:genetics*; D007527:Isoenzymes / Q000235:genetics*; D007685:Kidney Tubules, Collecting / Q000502:physiology; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D014018:Tissue Distribution; D024363:Transcription Initiation Site
D018076:DNA, Complementary; D007527:Isoenzymes; D012333:RNA, Messenger; D017506:H(+)-K(+)-Exchanging ATPase