Characterization of the chicken inward rectifier K+ channel IRK1/Kir2.1 gene.

Hideki Mutai, University of Pennsylvania School of Medicine
Lawrence C. Kenyon, Thomas Jefferson University
Emily Locke, University of Pennsylvania School of Medicine
Nami Kikuchi, University of Pennsylvania School of Medicine
John Carl Oberholtzer, University of Pennsylvania School of Medicine

Document Type Article

This article has been peer reviewed. It is the author’s final published version in BMC genomics

Volume 5, November 2004, Article number 90.

The published version is available at DOI: 10.1186/1471-2164-5-90. Copyright © Mutai et al.


BACKGROUND: Inward rectifier potassium channels (IRK) contribute to the normal function of skeletal and cardiac muscle cells. The chick inward rectifier K+ channel cIRK1/Kir2.1 is expressed in skeletal muscle, heart, brain, but not in liver; a distribution similar but not identical to that of mouse Kir2.1. We set out to explore regulatory domains of the cIRK1 promoter that enhance or inhibit expression of the gene in different cell types.

RESULTS: We cloned and characterized the 5'-flanking region of cIRK1. cIRK1 contains two exons with splice sites in the 5'-untranslated region, a structure similar to mouse and human orthologs. cIRK1 has multiple transcription initiation sites, a feature also seen in mouse. However, while the chicken and mouse promoter regions share many regulatory motifs, cIRK1 possesses a GC-richer promoter and a putative TATA box, which appears to positively regulate gene expression. We report here the identification of several candidate cell/tissue specific cIRK1 regulatory domains by comparing promoter activities in expressing (Qm7) and non-expressing (DF1) cells using in vitro transcription assays.

CONCLUSION: While multiple transcription initiation sites and the combinatorial function of several domains in activating cIRK1 expression are similar to those seen in mKir2.1, the cIRK1 promoter differs by the presence of a putative TATA box. In addition, several domains that regulate the gene's expression differentially in muscle (Qm7) and fibroblast cells (DF1) were identified. These results provide fundamental data to analyze cIRK1 transcriptional mechanisms. The control elements identified here may provide clues to the tissue-specific expression of this K+ channel.