Summary
Multidrug resistance-associated protein (MRP) and the canalicular multispecific organic anion transporter (cMOAT) are organic anion pumps that have been linked to cytotoxic drug resistance. We previously reported the isolation of three human MRP/cMOAT-related transporters, MOAT-B (MRP4), MOAT-C (MRP5) and MOAT-D (MRP3). In the present study we describe the fourth MRP/cMOAT-related transporter. We analysed ARA, a human cDNA reported to encode a 453 residue MRP-related transporter, and found that it represents a fused transcript composed of MRP sequences and partial sequences of a novel transporter. The complete coding sequence of this novel transporter, which we designated MOAT-E, was isolated. MOAT-E encodes a 1503 residue transporter that is most closely related to MRP (45%), MOAT-D (44%) and cMOAT (39%), both in terms of amino acid identity and sharing a common topology in which ~ 17 transmembrane spanning helices are distributed within three membrane spanning domains. RNA blot analysis indicated that MOAT-E expression is restricted to kidney and liver. These observations suggest that MOAT-E may function as an organic anion transporter involved in cellular detoxification and possibly in the hepatobiliary and renal excretion of xenobiotics and/or endogenous metabolites. Isolation of MOAT-E helps to define the MRP/cMOAT subfamily of transporters.
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References
Allegra, C. J. & Grem, J. L. (1997). Antimetabolites. In Cancer, Principles and Practice of Oncology, DeVita VT Jr, Hellman S, Rosenberg SA Lippincott-Raven: Philadelphia
Allikmets, R., Gerrard, B., Hutchinson, A. & Dean, M. (1996). Characterization of the human ABC superfamily: isolation and mapping of 21 new genes using the expressed sequence tags database. Hum Mol Genet 5: 1649–1655.
Bakos, E., Hegedus, T., Hollo, Z., Welker, E., Tusnady, G. E., Zaman, G. J., Flens, M. J., Varadi, A. & Sarkadi, B. (1996). Membrane topology and glycosylation of the human multidrug resistance-associated protein. J Biol Chem 271: 12322–12326.
Belinsky, M. G., Bain, L. J., Balsara, B. B., Testa, J. R. & Kruh, G. D. (1998). Characterization of MOAT-C and MOAT-D, new members of the MRP/cMOAT subfamily of transporter proteins. J Natl Cancer Inst 90: 1735–1741.
Breuninger, L. M., Paul, S., Gaughan, K., Miki, T., Chan, A., Aaronson, S. A. & Kruh, G. D. (1995). Expression of multidrug resistance-associated protein in NIH/3T3 cells confers multidrug resistance associated with increased drug efflux and altered intracellular drug distribution. Cancer Res 55: 5342–5347.
Buchler, M., Konig, J., Brom, M., Kartenbeck, J., Spring, H., Horie, T. & Keppler, D. (1996). cDNA cloning of the hepatocyte canalicular isoform of the multidrug resistance protein, cMrp, reveals a novel conjugate export pump deficient in hyperbilirubinemic mutant rats. J Biol Chem 271: 15091–15098.
Cole, S. P., Bhardwaj, G., Gerlach, J. H., Mackie, J. E., Grant, C. E., Almquist, K. C., Stewart, A. J., Kurz, E. U., Duncan, A. M. & Deeley, R. G. (1992). Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line [see comments]. Science 258: 1650–1654.
Cole, S. P., Sparks, K. E., Fraser, K., Loe, D. W., Grant, C. E., Wilson, G. M. & Deeley, R. G. (1994). Pharmacological characterization of multidrug resistant MRP-transfected human tumor cells. Cancer Res 54: 5902–5910.
Cui, Z., Hirata, D., Tsuchiya, E., Osada, H. & Miyakawa, T. (1996). The multidrug resistance-associated protein (MRP) subfamily (Yrs1/Yor1) of Saccharomyces cerevisiae is important for the tolerance to a broad range of organic anions. J Biol Chem 271: 14712–14716.
de Vries, M. H., Redegeld, F. A., Koster, A. S., Noordhoek, J., de Haan, J. G., Oude Elferink, R. P. & Jansen, P. L. (1989). Hepatic, intestinal and renal transport of 1-naphthol-beta-D-glucuronide in mutant rats with hereditary-conjugated hyperbilirubinemia. Naunyn Schmiedebergs Arch Pharmacol 340: 588–592.
Evers, R., Kool, M., van Deemter, L., Janssen, H., Calafat, J., Oomen, L. C., Paulusma, C. C., Oude Elferink, R. P., Baas, F., Schinkel, A. H. & Borst, P. (1998). Drug export activity of the human canalicular multispecific organic anion transporter in polarized kidney MDCK cells expressing cMOAT (MRP2) cDNA. J Clin Invest 101: 1310–1319.
Gottesman, M. M. & Pastan, I. (1993). Biochemistry of multidrug resistance mediated by the multidrug transporter. Annu Rev Biochem 62: 385–427.
Grant, C. E., Valdimarsson, G., Hipfner, D. R., Almquist, K. C., Cole, S. P. & Deeley, R. G. (1994). Overexpression of multidrug resistance-associated protein (MRP) increases resistance to natural product drugs. Cancer Res 54: 357–361.
Higgins, C. F. (1992). ABC transporters: from microorganisms to man. Annu Rev Cell Biol 8: 67–113.
Hipfner, D. R., Almquist, K. C., Leslie, E. M., Gerlach, J. H., Grant, C. E., Deeley, R. G. & Cole, S. P. (1997). Membrane topology of the multidrug resistance protein (MRP). A study of glycosylation-site mutants reveals an extracytosolic NH2 terminus. J Biol Chem 272: 23623–23630.
Huber, M., Guhlmann, A., Jansen, P. L. & Keppler, D. (1987). Hereditary defect of hepatobiliary cysteinyl leukotriene elimination in mutant rats with defective hepatic anion excretion. Hepatology 7: 224–228.
Ito, K., Suzuki, H., Hirohashi, T., Kume, K., Shimizu, T. & Sugiyama, Y. (1998). Functional analysis of a canalicular multispecific organic anion transporter cloned from rat liver. J Biol Chem 273: 1684–1688.
Jansen, P. L., Peters, W. H. & Lamers, W. H. (1985). Hereditary chronic conjugated hyperbilirubinemia in mutant rats caused by defective hepatic anion transport. Hepatology 5: 573–579.
Jedlitschky, G., Leier, I., Buchholz, U., Barnouin, K., Kurz, G. & Keppler, D. (1996). Transport of glutathione, glucuronate, and sulfate conjugates by the MRP gene-encoded conjugate export pump. Cancer Res 56: 988–994.
Kast, C. & Gros, P. (1997). Topology mapping of the amino-terminal half of multidrug resistance-associated protein by epitope insertion and immunofluorescence. J Biol Chem 272: 26479–26487.
Koike, K., Kawabe, T., Tanaka, T., Toh, S., Uchiumi, T., Wada, M., Akiyama, S., Ono, M. & Kuwano, M. (1997). A canalicular multispecific organic anion transporter (cMOAT) antisense cDNA enhances drug sensitivity in human hepatic cancer cells. Cancer Res 57: 5475–5479.
Kool, M., de Haas, M., Scheffer, G. L., Scheper, R. J., van Eijk, M. J., Juijn, J. A., Baas, F. & Borst, P. (1997). Analysis of expression of cMOAT (MRP2), MRP3, MRP4, and MRP5, homologues of the multidrug resistance-associated protein gene (MRP1), in human cancer cell lines. Cancer Res 57: 3537–3547.
Kruh, G. D., Chan, A., Myers, K., Gaughan, K., Miki, T. & Aaronson, S. A. (1994). Expression complementary DNA library transfer establishes mrp as a multidrug resistance gene. Cancer Res 54: 1649–1652.
Kruh, G. D., Gaughan, K. T., Godwin, A. & Chan, A. (1995). Expression pattern of MRP in human tissues and adult solid tumor cell lines. J Natl Cancer Inst 87: 1256–1258.
Kuss, B. J., O’Neill, G. M., Eyre, H., Doggett, N. A., Callen, D. F. & Davey, R. A. (1998). ARA, a novel ABC transporter, is located at 16p13.1, is deleted in inv(16) leukemias, and is shown to be expressed in primitive hematopoietic precursors. Genomics 51: 455–458.
Lee, K., Belinsky, M. G., Bell, D. W., Testa, J. R. & Kruh, G. D. (1998). Isolation of MOAT-B, a widely expressed multidrug resistance-associated protein/canalicular multispecific organic anion transporter-related transporter. Cancer Res 58: 2741–2747.
Leier, I., Jedlitschky, G., Buchholz, U., Cole, S. P., Deeley, R. G. & Keppler, D. (1994). The MRP gene encodes an ATP-dependent export pump for leukotriene C4 and structurally related conjugates. J Biol Chem 269: 27807–27810.
Leier, I., Jedlitschky, G., Buchholz, U., Center, M., Cole, S. P., Deeley, R. G. & Keppler, D. (1996). ATP-dependent glutathione disulphide transport mediated by the MRP gene-encoded conjugate export pump. Biochem J 314: 433–437.
Li, Z. S., Szczypka, M., Lu, Y. P., Thiele, D. J. & Rea, P. A. (1996). The yeast cadmium factor protein (YCF1) is a vacuolar glutathione S-conjugate pump. J Biol Chem 271: 6509–6517.
Loe, D. W., Almquist, K. C., Cole, S. P. & Deeley, R. G. (1996). ATP-dependent 17 beta-estradiol 17-(beta-D-glucuronide) transport by multidrug resistance protein (MRP). Inhibition by cholestatic steroids. J Biol Chem 271: 9683–9689.
Longhurst, T. J., O’Neill, G. M., Harvie, R. M. & Davey, R. A. (1996). The anthracycline resistance-associated (ara) gene, a novel gene associated with multidrug resistance in a human leukaemia cell line. Br J Cancer 74: 1331–1335.
Madon, J., Eckhardt, U., Gerloff, T., Stieger, B. & Meier, P. J. (1997). Functional expression of the rat liver canalicular isoform of the multidrug resistance-associated protein. FEBS Lett 406: 75–78.
Masuda, M., I’izuka, Y., Yamazaki, M., Nishigaki, R., Kato, Y., Ni’inuma, K., Suzuki, H. & Sugiyama, Y. (1997). Methotrexate is excreted into the bile by canalicular multispecific organic anion transporter in rats. Cancer Res 57: 3506–3510.
Mayer, R., Kartenbeck, J., Buchler, M., Jedlitschky, G., Leier, I. & Keppler, D. (1995). Expression of the MRP gene-encoded conjugate export pump in liver and its selective absence from the canalicular membrane in transport-deficient mutant hepatocytes. J Cell Biol 131: 137–150.
Mikami, T., Nozaki, T., Tagaya, O., Hosokawa, S., Nakura, T., Mori, H. & Kondou, S. (1986). The characters of a new mutant in rats with hyperbilirubinuria syndrome. Congenital Anom 26: 250–251.
Muller, M., Meijer, C., Zaman, G. J., Borst, P., Scheper, R. J., Mulder, N. H., de Vries, E. G. & Jansen, P. L. (1994). Overexpression of the gene encoding the multidrug resistance-associated protein results in increased ATP-dependent glutathione S-conjugate transport. Proc Natl Acad Sci USA 91: 13033–13037.
Neill, G. M., Peters, G. B., Harvie, R. M., MacKenzie, H. B., Henness, S. & Davey, R. A. (1998). Amplification and expression of the ABC transporters ARA and MRP in a series of multidrug-resistant leukaemia cell sublines. Br J Cancer 77: 2076–2080.
Ouellette, M., Fase-Fowler, F. & Borst, P. (1990). The amplified H circle of methotrexate-resistant Leishmania tarentolae contains a novel P-glycoprotein gene. Embo J 9: 1027–1033.
Paulusma, C. C., Bosma, P. J., Zaman, G. J., Bakker, C. T., Otter, M., Scheffer, G. L., Scheper, R. J., Borst, P. & Oude Elferink, R. P. (1996). Congenital jaundice in rats with a mutation in a multidrug resistance-associated protein gene. Science 271: 1126–1128.
Persson, B. & Argos, P. (1994). Prediction of transmembrane segments in proteins utilising multiple sequence alignments. J Mol Biol 237: 182–192.
Remon, A. M. H., van Aubel, R. A., van Kuijck, M. A., Koenderink, J. B., Deen, P. M., van Os, C. H. & Russel, F. G. (1998). Adenosine triphosphate-dependent transport of anionic conjugates by the rabbit multidrug resistance-associated protein Mrp2 expressed in insect cells. Mol Pharmacol 53: 1062–1067.
Schaub, T. P., Kartenbeck, J., Konig, J., Vogel, O., Witzgall, R., Kriz, W. & Keppler, D. (1997). Expression of the conjugate export pump encoded by the mrp2 gene in the apical membrane of kidney proximal tubules. J Am Soc Nephrol 8: 1213–1221.
Taniguchi, K., Wada, M., Kohno, K., Nakamura, T., Kawabe, T., Kawakami, M., Kagotani, K., Okumura, K., Akiyama, S. & Kuwano, M. (1996). A human canalicular multispecific organic anion transporter (cMOAT) gene is overexpressed in cisplatin-resistant human cancer cell lines with decreased drug accumulation. Cancer Res 56: 4124–4129.
Tusnady, G. E., Bakos, E., Varadi, A. & Sarkadi, B. (1997). Membrane topology distinguishes a subfamily of the ATP-binding cassette (ABC) transporters. FEBS Lett 402: 1–3.
Zaman, G. J., Flens, M. J., van Leusden, M. R., de Haas, M., Mulder, H. S., Lankelma, J., Pinedo, H. M., Scheper, R. J., Baas, F., Broxterman, H. J. & Borst, P. (1994). The human multidrug resistance-associated protein MRP is a plasma membrane drug-efflux pump. Proc Natl Acad Sci USA 91: 8822–8826.
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Belinsky, M., Kruh, G. MOAT-E (ARA) is a full-length MRP/cMOAT subfamily transporter expressed in kidney and liver. Br J Cancer 80, 1342–1349 (1999). https://doi.org/10.1038/sj.bjc.6690527
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DOI: https://doi.org/10.1038/sj.bjc.6690527
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