Cloning and functional expression of human aquaporin8 cDNA and analysis of its gene.
The cDNA and the gene of human aquaporin8 (AQP8) were cloned from human testis cDNA and a genomic library, respectively. The AQP8 cDNA encodes 261 amino acids. The identity of the amino acid sequence to other aquaporins is highest with a plant water channel, gamma-TIP (40.4%), while AQP2 and AQP3 are 28.9 and 29.5% identical to human AQP8, respectively. The human AQP8 is only 74.9% identical to rat AQP8 and 76.0% identical to mouse AQP8. In Northern blot analysis, approximately 1.35-kb human AQP8 mRNA was expressed in pancreas and colon, but not in other tissues. Absence of human AQP8 in testis is noteworthy as rat AQP8 was abundantly expressed in testis. The expression of human AQP8 cRNA in Xenopus oocytes increased osmotic water permeability by 10-fold. AQP8 was not permeable to urea nor to glycerol. The AQP8 gene has five introns, and the locations of exon-intron boundaries were different from those of the other mammalian aquaporins, suggesting its separate phylogenetic origin.
Koyama
N
N
Second Department of Internal Medicine, Tokyo Medical and Dental University, Tokyo, 1-5-45 Yushima, Bunkyo-ku, 113-8519, Japan.
Ishibashi
K
K
Kuwahara
M
M
Inase
N
N
Ichioka
M
M
Sasaki
S
S
Marumo
F
F
D000595:Amino Acid Sequence; D000818:Animals; D020346:Aquaporins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015152:Blotting, Northern; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005091:Exons / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007438:Introns / Q000235:genetics; D007473:Ion Channels; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity; D051381:Rats; D016415:Sequence Alignment; D013737:Testis / Q000378:metabolism*
D020346:Aquaporins; D018076:DNA, Complementary; D007473:Ion Channels; C108330:aquaporin 8