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Am J Physiol Cell Physiol 276: C585-C592, 1999;
0363-6143/99 $5.00
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Vol. 276, Issue 3, C585-C592, March 1999

Splicing of a retained intron within ROMK K+ channel RNA generates a novel set of isoforms in rat kidney

A. H. Beesley, B. Ortega, and S. J. White

Laboratory for Membrane Protein Function, Department of Biomedical Science, University of Sheffield, Sheffield S10 2TN, United Kingdom

The renal outer medulla K+ channel (ROMK) family of K+ channels may constitute a major pathway for K+ secretion in the distal nephron. To date, four main isoforms of this gene have been identified in the rat that differ only in their NH2-terminal amino acids and that share a common "core exon" that determines the remaining protein sequence. Using RT-PCR, we have identified a new set of ROMK isoforms in rat kidney that are generated by the deletion of a region within the ROMK core sequence that is identifiable as a typical mammalian intron. This splicing event was shown to be reproducible in vitro by detection of deleted ROMK mRNA in Madin-Darby canine kidney (MDCK) cells stably transfected with the gene for ROMK2. Translation of the deletion variant of ROMK2 was confirmed in vitro and visualized in MDCK cells following transient transfection with an enhanced green fluorescent protein tag. The deletion in this core region is predicted to generate hydrophilic proteins that are approximately one-third of the size of native ROMK and lack membrane-spanning domains.

reverse transcriptase-polymerase chain reaction; alternative splicing; inward rectifier; accessory proteins; ion channel; renal outer medulla potassium channel


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