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. 1995 Jan 1;305(Pt 1):81–85. doi: 10.1042/bj3050081

Cloning and functional expression of a Na(+)-dependent phosphate co-transporter from human kidney: cDNA cloning and functional expression.

K Miyamoto 1, S Tatsumi 1, T Sonoda 1, H Yamamoto 1, H Minami 1, Y Taketani 1, E Takeda 1
PMCID: PMC1136432  PMID: 7826357

Abstract

A cDNA clone encoding a protein 69% identical in amino acid sequence with that of the Na/P(i) co-transporter NaP(i)-1 was isolated from a human kidney cDNA library. The DNA sequence was identical with that of NPT-1 cDNA published by Chong, Kristjansson, Zoghbi and Hughe (1993) (Genomics, 18, 355-359). In the present study, we have characterized the function of the encoded protein and the tissue distribution of its mRNA. Injection of RNA transcribed from NPT-1 into Xenopus oocytes resulted in expression of Na/P(i) co-transport activity showing a high affinity for P(i) transport (Km 0.29 mM). Kinetic characterization ([P(i)], [Na+]) demonstrated that the expressed transport activity has properties similar to those displayed by oocytes injected with human kidney poly(A)+ RNA. Northern blotting demonstrated that NPT-1 mRNA is expressed in renal cortex, liver and brain but not in other tissues. Hybrid depletion with antisense oligonucleotides to NaP(i)-3 and NPT-1 completely inhibited poly(A)+ RNA-induced Na(+)-dependent P(i) uptake in oocytes. These findings indicate that two high-affinity Na/P(i) cotransporters (NaP(i)-3 and NPT-1) are present in human kidney cortex.

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Selected References

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  1. Biber J., Custer M., Werner A., Kaissling B., Murer H. Localization of NaPi-1, a Na/Pi cotransporter, in rabbit kidney proximal tubules. II. Localization by immunohistochemistry. Pflugers Arch. 1993 Aug;424(3-4):210–215. doi: 10.1007/BF00384344. [DOI] [PubMed] [Google Scholar]
  2. Bindels R. J., van den Broek L. A., van Os C. H. Effect of pH on the kinetics of Na+-dependent phosphate transport in rat renal brush-border membranes. Biochim Biophys Acta. 1987 Feb 12;897(1):83–92. doi: 10.1016/0005-2736(87)90317-8. [DOI] [PubMed] [Google Scholar]
  3. Brunette M. G., Chan M., Maag U., Béliveau R. Phosphate uptake by superficial and deep nephron brush border membranes. Effect of the dietary phosphate and parathyroid hormone. Pflugers Arch. 1984 Apr;400(4):356–362. doi: 10.1007/BF00587532. [DOI] [PubMed] [Google Scholar]
  4. Brunette M. G., Chan M., Maag U., Béliveau R. Phosphate uptake by superficial and deep nephron brush border membranes. Effect of the dietary phosphate and parathyroid hormone. Pflugers Arch. 1984 Apr;400(4):356–362. doi: 10.1007/BF00587532. [DOI] [PubMed] [Google Scholar]
  5. Chong S. S., Kristjansson K., Zoghbi H. Y., Hughes M. R. Molecular cloning of the cDNA encoding a human renal sodium phosphate transport protein and its assignment to chromosome 6p21.3-p23. Genomics. 1993 Nov;18(2):355–359. doi: 10.1006/geno.1993.1476. [DOI] [PubMed] [Google Scholar]
  6. Contreras R., Cheroutre H., Degrave W., Fiers W. Simple, efficient in vitro synthesis of capped RNA useful for direct expression of cloned eukaryotic genes. Nucleic Acids Res. 1982 Oct 25;10(20):6353–6362. doi: 10.1093/nar/10.20.6353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Custer M., Meier F., Schlatter E., Greger R., Garcia-Perez A., Biber J., Murer H. Localization of NaPi-1, a Na-Pi cotransporter, in rabbit kidney proximal tubules. I. mRNA localization by reverse transcription/polymerase chain reaction. Pflugers Arch. 1993 Aug;424(3-4):203–209. doi: 10.1007/BF00384343. [DOI] [PubMed] [Google Scholar]
  8. Delisle M. C., Boyer C., Vachon V., Giroux S., Béliveau R. Immunodetection and characterization of proteins implicated in renal sodium/phosphate cotransport. Biochim Biophys Acta. 1994 Mar 23;1190(2):289–296. doi: 10.1016/0005-2736(94)90086-8. [DOI] [PubMed] [Google Scholar]
  9. Ghishan F. K., Rebeiz R., Honda T., Nakagawa N. Characterization and expression of a novel Na(+)-inorganic phosphate transporter at the liver plasma membrane of the rat. Gastroenterology. 1993 Aug;105(2):519–526. doi: 10.1016/0016-5085(93)90729-v. [DOI] [PubMed] [Google Scholar]
  10. Kos C. H., Tihy F., Econs M. J., Murer H., Lemieux N., Tenenhouse H. S. Localization of a renal sodium-phosphate cotransporter gene to human chromosome 5q35. Genomics. 1994 Jan 1;19(1):176–177. doi: 10.1006/geno.1994.1034. [DOI] [PubMed] [Google Scholar]
  11. Magagnin S., Werner A., Markovich D., Sorribas V., Stange G., Biber J., Murer H. Expression cloning of human and rat renal cortex Na/Pi cotransport. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):5979–5983. doi: 10.1073/pnas.90.13.5979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Miyamoto K., Hase K., Taketani Y., Minami H., Oka T., Nakabou Y., Hagihira H. Developmental changes in intestinal glucose transporter mRNA levels. Biochem Biophys Res Commun. 1992 Mar 16;183(2):626–631. doi: 10.1016/0006-291x(92)90528-s. [DOI] [PubMed] [Google Scholar]
  13. Miyamoto K., Hase K., Taketani Y., Minami H., Oka T., Nakabou Y., Hagihira H. Diabetes and glucose transporter gene expression in rat small intestine. Biochem Biophys Res Commun. 1991 Dec 31;181(3):1110–1117. doi: 10.1016/0006-291x(91)92053-m. [DOI] [PubMed] [Google Scholar]
  14. Scriver C. R., Tenenhouse H. S. X-linked hypophosphataemia: a homologous phenotype in humans and mice with unusual organ-specific gene dosage. J Inherit Metab Dis. 1992;15(4):610–624. doi: 10.1007/BF01799618. [DOI] [PubMed] [Google Scholar]
  15. Sorribas V., Markovich D., Hayes G., Stange G., Forgo J., Biber J., Murer H. Cloning of a Na/Pi cotransporter from opossum kidney cells. J Biol Chem. 1994 Mar 4;269(9):6615–6621. [PubMed] [Google Scholar]
  16. Surin B. P., Jans D. A., Fimmel A. L., Shaw D. C., Cox G. B., Rosenberg H. Structural gene for the phosphate-repressible phosphate-binding protein of Escherichia coli has its own promoter: complete nucleotide sequence of the phoS gene. J Bacteriol. 1984 Mar;157(3):772–778. doi: 10.1128/jb.157.3.772-778.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Tenenhouse H. S., Henry H. L. Protein kinase activity and protein kinase inhibitor in mouse kidney: effect of the X-linked Hyp mutation and vitamin D status. Endocrinology. 1985 Nov;117(5):1719–1726. doi: 10.1210/endo-117-5-1719. [DOI] [PubMed] [Google Scholar]
  18. Tenenhouse H. S., Klugerman A. H., Neal J. L. Effect of phosphonoformic acid, dietary phosphate and the Hyp mutation on kinetically distinct phosphate transport processes in mouse kidney. Biochim Biophys Acta. 1989 Sep 4;984(2):207–213. doi: 10.1016/0005-2736(89)90218-6. [DOI] [PubMed] [Google Scholar]
  19. Tenenhouse H. S., Werner A., Biber J., Ma S., Martel J., Roy S., Murer H. Renal Na(+)-phosphate cotransport in murine X-linked hypophosphatemic rickets. Molecular characterization. J Clin Invest. 1994 Feb;93(2):671–676. doi: 10.1172/JCI117019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Walker J. J., Yan T. S., Quamme G. A. Presence of multiple sodium-dependent phosphate transport processes in proximal brush-border membrane. Am J Physiol. 1987 Feb;252(2 Pt 2):F226–F231. doi: 10.1152/ajprenal.1987.252.2.F226. [DOI] [PubMed] [Google Scholar]
  21. Wehrle J. P., Pedersen P. L. Characteristics of phosphate uptake by Ehrlich ascites tumor cells. J Biol Chem. 1982 Aug 25;257(16):9698–9703. [PubMed] [Google Scholar]
  22. Werner A., Kempson S. A., Biber J., Murer H. Increase of Na/Pi-cotransport encoding mRNA in response to low Pi diet in rat kidney cortex. J Biol Chem. 1994 Mar 4;269(9):6637–6639. [PubMed] [Google Scholar]
  23. Werner A., Moore M. L., Mantei N., Biber J., Semenza G., Murer H. Cloning and expression of cDNA for a Na/Pi cotransport system of kidney cortex. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9608–9612. doi: 10.1073/pnas.88.21.9608. [DOI] [PMC free article] [PubMed] [Google Scholar]

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