Cloning of a human nucleoside transporter implicated in the Cellular uptake of adenosine and chemotherapeutic drugs

Abstract

In most mammalian Cells nucleoside uptake occurs primarily via broad-specificity, es (e, equilibrative; s, sensitive to NBMPR inhibition) transporters that are potentlyinhibited by nitrobenzylthioinosine (NBMPR)1. These transporters are essential for nucleotide synthesis by salvage pathways in hemopoietic and other Cells that lack de novo pathways and are the route of Cellular uptake for many cytotoxic nucleosides used in cancer and viral chemotherapy1. They play an important role in adenosinemediated regulation of many physiological processes, including neurotransmission and platelet aggregation, and are a target for coronary vasodilator drugs. We have previously reported the purification of the prototypic es transporter from human erythrocytes2 and have shown that this glycoprotein of apparent M, 55,000 is immunologically related to nucleoside transporters from several other species and tissues, including human placenta3,4. Here we report the isolation of a human placental cDNA encoding a 456-residue glycoprotein with functional characteristics typical of an es-type transporter. It is predicted to possess 11 membrane-spanning regions and is homologous to several proteins of unknown function in yeast, nematodes, plants and mammals. Because of its central role in the uptake both of adenosine and of chemotherapeutic nucleosides, study of this protein should not only provide insights into the physiological roles of nucleoside transport but also open the way to improved therapies.

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References

  1. 1

    Cass, C.E. Nucleoside transport. in Drug Transport in Antimicrobial and Anticancer Chemotherapy (ed Georgopapadakou, N. H.) 403–451 (Marcel Dekker, New York, ( 1995).

  2. 2

    Kwong, F.Y.P. et al. Purification of the human erythrocyte nudeoside transporter by immunoaffinity chromatography. Biochem. J. 255, 243–249 ( 1988).

  3. 3

    Kwong, F.Y.P. et al. Mammalian nitrobenzylthioinosine-sensitive nudeoside transport proteins: Immunological evidence that transporters differing in size and inhibitor-sensitivity share sequence homology. J. Biol. Chem. 267, 21954–21960 ( 1992).

  4. 4

    Barros, L.F. et al. Immunolocalisation of nudeoside transporters in human placental trophoblast and endothelial Cells: Evidence for multiple transporter isoforms. Pfluegen Arch.Eur. J. Physiol. 429, 394–399 ( 1995).

  5. 5

    Kwong, F.Y.P. et al. Enzymic cleavage as a probe of the molecular structures of mammalian equilibrative nudeoside transporters. J. Biol. Chem. 268, 22127–22134 ( 1993).

  6. 6

    Huang, Q.Q. et al. Cloning and functional expression of a complementary DNA encoding a mammalian nudeosidetransport protein. J. Biol. Chem. 269, 17757–17760 ( 1994).

  7. 7

    Che, M., Ortiz, D.F. & Arias, I.M. Primary structure and functional expression of a cDNA encoding the bile canalicular, purine-specific Na+-nudeoside co-transporter. J. Biol. Chem. 270, 13596–13599 ( 1995).

  8. 8

    Craig, J.E., Zhang, Y. & Gallagher, M.P. Cloning of the nupC gene of Escherichia coli encoding a nucleoside transport system, and identification of an adjacent insertion element, IS 186. Mol. Microbiol. 11, 1159–1168 ( 1994).

  9. 9

    Westh-Hansen, S.E., Jensen, N. & Munch-Petersen, A. Studies on the sequence and structure of the Escherichiacoli K-12 nupG gene, encoding a nucleoside-transport system. Eur. J. Biochem. 168, 385–391 ( 1987).

  10. 10

    Ouellette, B.F.F. et al. Sequencing of chromosome I from Saccharomyces cerevisiae:. Analysis of a 32 kb region between the LTE1 and SPO7 genes. Genome 36, 32–42 ( 1993).

  11. 11

    Williams, J.B. & Lanahan, A.A. A mammalian delayed-early response gene encodes HNP36, a novel, conserved nucleolar protein. Biochem. Biophys. Res. Commun. 213, 325–333 ( 1995).

  12. 12

    Huang, Q.Q., Harvey, CM., Paterson, A.R., Cass, C.E. & Young, J.D. Functional expression of Na+-dependent nucleoside transport systems of rat intestine in isolated oocytes of. Xenopus laevis. J. Biol. Chem. 268, 20613–20619 ( 1993).

  13. 13

    Young, J.D. & Jarvis, S.M. Nucleoside transport in animal Cells. Biosci. Rep. 3, 309–322 ( 1983).

  14. 14

    Bryson, H.M. & Sorkin, E.M., A review of its pharmacodynamic and pharmacokinetic properties and therapeutic potential in haematological malignancies. Drugs 46, 872–894 ( 1993).

  15. 15

    Rustum, Y.M. & Raymakers, R.A.P. 1-β-Arabinofuranosylcytosine in therapy of leukaemia: Predinical and clinical overview. Pharmacol. Ther. 56, 307–321 ( 1992).

  16. 16

    Ross, S.R., McTavish, D. & Faulds, D. Fludarabine: A review of its pharmacological properties and therapeutic potential in malignancy. Drugs 45, 737–759 ( 1993).

  17. 17

    Kaye, S.B., Gemcitabine: Current status of phase I and II trials. J. Clin. Oncol. 12, 1527–1531 ( 1994).

  18. 18

    Mizushima, S. & Nagata, S. pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res. 18, 5322 ( 1990).

  19. 19

    Kozak, M. The scanning model for translation: An update. J. Cell Biol. 108, 229–241 ( 1989).

  20. 20

    Hofmann, K. & Stoffel, W., TMbase—A database of membrane spanning protein segments. Biol. Chem. Hoppe-Seyler 347, 166 ( 1993).

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Griffiths, M., Beaumont, N., Yao, S. et al. Cloning of a human nucleoside transporter implicated in the Cellular uptake of adenosine and chemotherapeutic drugs. Nat Med 3, 89–93 (1997). https://doi.org/10.1038/nm0197-89

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