Identification of a novel K-channel gene (KC22) that is highly expressed in distal tubule of rabbit kidney.
The Shaker gene family encodes voltage-gated K channels. Five partial-length Shaker-like cDNAs (KC2, 4, 10, 19, and 22) were previously isolated from rabbit kidney using polymerase chain reaction (PCR) [G. V. Desir, E. Hamlin, A.H. Puente, R.F. Reilly, F. Hiledebrandt, and P. Igarashi. Am. J. Physiol. 262 (Renal Fluid Electrolyte Physiol. 31): F151-F157, 1992]. We now report the cloning of another Shaker-like cDNA (KC6) from rabbit kidney and the identification of one isoform that is highly expressed in rabbit distal tubule cells grown in culture. A partial-length cDNA (859 bp) for KC6 was isolated by PCR amplification of rabbit kidney cDNA using Shaker-specific degenerate primers. KC6 was most similar to the rat brain clone RBK2 (77% amino acid identity) and to the rabbit clone KC19 (78% amino acid identity). Transcript levels for KC2, 4, 6, 10, 19, and 22 were quantified using the ribonuclease protection assay. Transcripts for all six isoforms were detected in renal tissues. KC22 was the most abundant isoform in kidney cortex and medulla (20- to 40-fold greater than the other isoforms). Furthermore, KC22 expression levels were fivefold higher in primary cultures of rabbit distal convoluted tubules and connecting tubules than in whole kidney cortex. Although the partial-length sequence for KC22 represents the most conserved regions in the Shaker gene family it only has 35-88% amino acid identity with other Shaker channels, suggesting that KC22 represents a novel isoform. In contrast, KC4 and KC19 (less abundant in kidney than KC22) are highly homologous to the rat brain clones RBK1 and RBK2, respectively (97% amino acid identity).(ABSTRACT TRUNCATED AT 250 WORDS)
Desir
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Department of Medicine, Yale University School of Medicine, New Haven, Connecticut 06510.
Velázquez
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D000818:Animals; D001483:Base Sequence; D004330:Drosophila / Q000235:genetics; D015870:Gene Expression; D007685:Kidney Tubules, Collecting / Q000378:metabolism / Q000502:physiology; D007686:Kidney Tubules, Distal / Q000378:metabolism / Q000502:physiology*; D008969:Molecular Sequence Data; D009399:Nephrons / Q000378:metabolism; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D015221:Potassium Channels / Q000235:genetics*; D012313:RNA / Q000378:metabolism; D011817:Rabbits; D017385:Sequence Homology; D014158:Transcription, Genetic
D015345:Oligonucleotide Probes; D015221:Potassium Channels; D012313:RNA