title: cDNA cloning and gene structure of a novel water channel expressed exclusively in human kidney: evidence for a gene cluster of aquaporins at chromosome locus 12q13.
abstract: A 1.8-kb cDNA clone (designed hKID, gene symbol AQP2L) with homology to the aquaporins was isolated from a human kidney cDNA library. The longest open reading frame of 846 bp encoded a 282-amino-acid hydrophobic protein that contained the conserved NPA motifs of MIP family members. Cell-free translation produced a nonglycosylated protein migrating at 29 kDa. Amino acid alignment showed the greatest homology of hKID to human MIP (48% identity) and AQP-2 (52%), with lesser homology to human MIWC (AQP-4, 34%), CHIP28 (AQP-1, 38%), and GLIP (AQP-3, 22%). Northern blot analysis revealed a 2.2-kb transcript expressed only in human kidney. PCR/Southern blot analysis of human kidney cDNA using primers flanking the hKID coding sequence revealed expression of a full-length mRNA and short transcripts with partial exon 1 and partial exon 4 deletions. Expression of hKID cRNA in Xenopus oocytes did not increase glycerol or urea permeability, but increased osmotic water permeability from (2.8 +/- 0.5) x 10(-4) to (7.4 +/- 0.7) x 10(-4) cm/s (10 degrees C) in a mercurial-sensitive manner. Sequence comparison of hKID cDNA with a cloned 21-kb genomic DNA indicated three introns (lengths 0.7, 0.25, and 0.4 kb) separating four exons with boundaries at amino acids 121, 174, and 201. The hKID promoter was identified and contained TATA, SP1, E-box, and AP1 and AP2 elements; primer extension revealed hKID transcription initiation 654 bp upstream from the translational initiation site. Genomic Southern blot indicated a single-copy hKID gene. PCR analysis of a human/rodent somatic hybrid panel localized the hKID gene to chromosome 12. Chromosomal fluorescence in situ hybridization mapped the hKID (AQP2L) gene to chromosome locus 12q13, the same location as the AQP. 2 and MIP genes. The high sequence homology, similar genomic structure, and identical chromosomal loci of hKID, MIP, and AQP-2 suggest a MIP family gene cluster at chromosome locus 12q13. Further work is needed to establish the physiological significance of hKID.
authors.affiliation: Department of Medicine, University of California, San Francisco 94143-0521, USA.
authors.forename: T
authors.initials: T
authors.lastname: Ma
authors.forename: B
authors.initials: B
authors.lastname: Yang
authors.forename: W L
authors.initials: WL
authors.lastname: Kuo
authors.forename: A S
authors.initials: AS
authors.lastname: Verkman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D001483:Base Sequence; D002881:Chromosomes, Human, Pair 12; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary; D015870:Gene Expression; D006801:Humans; D007473:Ion Channels / Q000235:genetics*; D007668:Kidney / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D005810:Multigene Family; D017386:Sequence Homology, Amino Acid; D014018:Tissue Distribution; D014867:Water; D014981:Xenopus
chemical_list: C496023:AQP2 protein, human; C102226:AQP6 protein, human; C496024:Aqp2 protein, mouse; D051399:Aquaporin 2; D051403:Aquaporin 6; D020346:Aquaporins; D018076:DNA, Complementary; D007473:Ion Channels; D014867:Water