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. 1997 Sep 15;326(Pt 3):897–902. doi: 10.1042/bj3260897

Two forms of Wilson disease protein produced by alternative splicing are localized in distinct cellular compartments.

X L Yang 1, N Miura 1, Y Kawarada 1, K Terada 1, K Petrukhin 1, T Gilliam 1, T Sugiyama 1
PMCID: PMC1218748  PMID: 9307043

Abstract

Copper is an essential trace element in prokaryotes and eukaryotes and is strictly regulated by biological mechanisms. Menkes and Wilson diseases are human disorders that arise from disruption of the normal process of copper export from the cytosol to the extracellular environment. Recently a gene for Wilson disease (WD)(also named the ATP7B gene) was cloned. This gene encodes a copper transporter of the P-type ATPase. We prepared monoclonal and polyclonal anti-(WD protein) antibodies and characterized the full-length WD protein as well as a shorter form that is produced by alternative splicing in the human brain. We found that the WD protein is localized mainly in the Golgi apparatus, whereas the shorter form is present in the cytosol. These results suggest that the alternative WD proteins act as key regulators of copper metabolism, perhaps by performing distinct roles in the intracellular transport and export of copper.

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Selected References

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  1. Amoureux M. C., Wurch T., Pauwels P. J. Modulation of metallothionein-III mRNA content and growth rate of rat C6-glial cells by transfection with human 5-HT1D receptor genes. Biochem Biophys Res Commun. 1995 Sep 14;214(2):639–645. doi: 10.1006/bbrc.1995.2334. [DOI] [PubMed] [Google Scholar]
  2. Bingham M. J., Burchell A., McArdle H. J. Identification of an ATP-dependent copper transport system in endoplasmic reticulum vesicles isolated from rat liver. J Physiol. 1995 Feb 1;482(Pt 3):583–587. doi: 10.1113/jphysiol.1995.sp020542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bull P. C., Cox D. W. Wilson disease and Menkes disease: new handles on heavy-metal transport. Trends Genet. 1994 Jul;10(7):246–252. doi: 10.1016/0168-9525(94)90172-4. [DOI] [PubMed] [Google Scholar]
  4. Bull P. C., Thomas G. R., Rommens J. M., Forbes J. R., Cox D. W. The Wilson disease gene is a putative copper transporting P-type ATPase similar to the Menkes gene. Nat Genet. 1993 Dec;5(4):327–337. doi: 10.1038/ng1293-327. [DOI] [PubMed] [Google Scholar]
  5. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Choudhuri S., Kramer K. K., Berman N. E., Dalton T. P., Andrews G. K., Klaassen C. D. Constitutive expression of metallothionein genes in mouse brain. Toxicol Appl Pharmacol. 1995 Mar;131(1):144–154. doi: 10.1006/taap.1995.1056. [DOI] [PubMed] [Google Scholar]
  7. Cousins R. J. Absorption, transport, and hepatic metabolism of copper and zinc: special reference to metallothionein and ceruloplasmin. Physiol Rev. 1985 Apr;65(2):238–309. doi: 10.1152/physrev.1985.65.2.238. [DOI] [PubMed] [Google Scholar]
  8. Dijkstra M., In 't Veld G., van den Berg G. J., Müller M., Kuipers F., Vonk R. J. Adenosine triphosphate-dependent copper transport in isolated rat liver plasma membranes. J Clin Invest. 1995 Jan;95(1):412–416. doi: 10.1172/JCI117670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hartmann H. A., Evenson M. A. Deficiency of copper can cause neuronal degeneration. Med Hypotheses. 1992 May;38(1):75–85. doi: 10.1016/0306-9877(92)90162-6. [DOI] [PubMed] [Google Scholar]
  10. Heuser J. Effects of cytoplasmic acidification on clathrin lattice morphology. J Cell Biol. 1989 Feb;108(2):401–411. doi: 10.1083/jcb.108.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Iyengar V., Brewer G. J., Dick R. D., Chung O. Y. Studies of cholecystokinin-stimulated biliary secretions reveal a high molecular weight copper-binding substance in normal subjects that is absent in patients with Wilson's disease. J Lab Clin Med. 1988 Mar;111(3):267–274. [PubMed] [Google Scholar]
  12. Masters B. A., Quaife C. J., Erickson J. C., Kelly E. J., Froelick G. J., Zambrowicz B. P., Brinster R. L., Palmiter R. D. Metallothionein III is expressed in neurons that sequester zinc in synaptic vesicles. J Neurosci. 1994 Oct;14(10):5844–5857. doi: 10.1523/JNEUROSCI.14-10-05844.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Miura N., Uchida T., Okada Y. HVJ (Sendai virus)-induced envelope fusion and cell fusion are blocked by monoclonal anti-HN protein antibody that does not inhibit hemagglutination activity of HVJ. Exp Cell Res. 1982 Oct;141(2):409–420. doi: 10.1016/0014-4827(82)90229-4. [DOI] [PubMed] [Google Scholar]
  14. Murata Y., Yamakawa E., Iizuka T., Kodama H., Abe T., Seki Y., Kodama M. Failure of copper incorporation into ceruloplasmin in the Golgi apparatus of LEC rat hepatocytes. Biochem Biophys Res Commun. 1995 Apr 6;209(1):349–355. doi: 10.1006/bbrc.1995.1510. [DOI] [PubMed] [Google Scholar]
  15. Niwa H., Yamamura K., Miyazaki J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene. 1991 Dec 15;108(2):193–199. doi: 10.1016/0378-1119(91)90434-d. [DOI] [PubMed] [Google Scholar]
  16. Petris M. J., Mercer J. F., Culvenor J. G., Lockhart P., Gleeson P. A., Camakaris J. Ligand-regulated transport of the Menkes copper P-type ATPase efflux pump from the Golgi apparatus to the plasma membrane: a novel mechanism of regulated trafficking. EMBO J. 1996 Nov 15;15(22):6084–6095. [PMC free article] [PubMed] [Google Scholar]
  17. Petrukhin K., Fischer S. G., Pirastu M., Tanzi R. E., Chernov I., Devoto M., Brzustowicz L. M., Cayanis E., Vitale E., Russo J. J. Mapping, cloning and genetic characterization of the region containing the Wilson disease gene. Nat Genet. 1993 Dec;5(4):338–343. doi: 10.1038/ng1293-338. [DOI] [PubMed] [Google Scholar]
  18. Petrukhin K., Lutsenko S., Chernov I., Ross B. M., Kaplan J. H., Gilliam T. C. Characterization of the Wilson disease gene encoding a P-type copper transporting ATPase: genomic organization, alternative splicing, and structure/function predictions. Hum Mol Genet. 1994 Sep;3(9):1647–1656. doi: 10.1093/hmg/3.9.1647. [DOI] [PubMed] [Google Scholar]
  19. Prohaska J. R., Wells W. W. Copper deficiency in the developing rat brain: a possible model for Menkes' steely-hair disease. J Neurochem. 1974 Jul;23(1):91–98. doi: 10.1111/j.1471-4159.1974.tb06920.x. [DOI] [PubMed] [Google Scholar]
  20. Samuels A. R., Freedman J. H., Bhargava M. M. Purification and characterization of a novel abundant protein in rat bile that binds azo dye metabolites and copper. Biochim Biophys Acta. 1983 Aug 23;759(1-2):23–31. doi: 10.1016/0304-4165(83)90184-8. [DOI] [PubMed] [Google Scholar]
  21. Sato M., Bremner I. Biliary excretion of metallothionein and a possible degradation product in rats injected with copper and zinc. Biochem J. 1984 Oct 15;223(2):475–479. doi: 10.1042/bj2230475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sato M., Gitlin J. D. Mechanisms of copper incorporation during the biosynthesis of human ceruloplasmin. J Biol Chem. 1991 Mar 15;266(8):5128–5134. [PubMed] [Google Scholar]
  23. Solioz M., Vulpe C. CPx-type ATPases: a class of P-type ATPases that pump heavy metals. Trends Biochem Sci. 1996 Jul;21(7):237–241. [PubMed] [Google Scholar]
  24. Sternlieb I., Van den Hamer C. J., Morell A. G., Alpert S., Gregoriadis G., Scheinberg I. H. Lysosomal defect of hepatic copper excretion in Wilson's disease (hepatolenticular degeneration). Gastroenterology. 1973 Jan;64(1):99–105. [PubMed] [Google Scholar]
  25. Storrie B., Madden E. A. Isolation of subcellular organelles. Methods Enzymol. 1990;182:203–225. doi: 10.1016/0076-6879(90)82018-w. [DOI] [PubMed] [Google Scholar]
  26. Tanzi R. E., Petrukhin K., Chernov I., Pellequer J. L., Wasco W., Ross B., Romano D. M., Parano E., Pavone L., Brzustowicz L. M. The Wilson disease gene is a copper transporting ATPase with homology to the Menkes disease gene. Nat Genet. 1993 Dec;5(4):344–350. doi: 10.1038/ng1293-344. [DOI] [PubMed] [Google Scholar]
  27. Tashima Y., Mizunuma H., Shima H., Kobayashi R., Hakeda Y., Maeda N., Kumegawa M. Effect of glucocorticoids on induction of fructose bisphosphatase and glucose-6-phosphatase in fetal mouse liver. J Biochem. 1984 Nov;96(5):1619–1624. doi: 10.1093/oxfordjournals.jbchem.a134992. [DOI] [PubMed] [Google Scholar]
  28. Terada K., Kawarada Y., Miura N., Yasui O., Koyama K., Sugiyama T. Copper incorporation into ceruloplasmin in rat livers. Biochim Biophys Acta. 1995 Jan 25;1270(1):58–62. doi: 10.1016/0925-4439(94)00072-x. [DOI] [PubMed] [Google Scholar]
  29. Terao T., Owen C. A., Jr Nature of copper compounds in liver supernate and bile of rats: studies with 67 Cu. Am J Physiol. 1973 Mar;224(3):682–686. doi: 10.1152/ajplegacy.1973.224.3.682. [DOI] [PubMed] [Google Scholar]
  30. Urayama O., Nagamune H., Nakao M., Hara Y. A monoclonal antibody against a native conformation of the porcine renal Na+/K(+)-ATPase alpha-subunit protein. Biochim Biophys Acta. 1990 Sep 3;1040(2):267–275. doi: 10.1016/0167-4838(90)90086-u. [DOI] [PubMed] [Google Scholar]
  31. WARREN P. J., EARL C. J., THOMPSON R. H. The distribution of copper in human brain. Brain. 1960 Dec;83:709–717. doi: 10.1093/brain/83.4.709. [DOI] [PubMed] [Google Scholar]
  32. Yamaguchi Y., Heiny M. E., Suzuki M., Gitlin J. D. Biochemical characterization and intracellular localization of the Menkes disease protein. Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):14030–14035. doi: 10.1073/pnas.93.24.14030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yuan D. S., Stearman R., Dancis A., Dunn T., Beeler T., Klausner R. D. The Menkes/Wilson disease gene homologue in yeast provides copper to a ceruloplasmin-like oxidase required for iron uptake. Proc Natl Acad Sci U S A. 1995 Mar 28;92(7):2632–2636. doi: 10.1073/pnas.92.7.2632. [DOI] [PMC free article] [PubMed] [Google Scholar]

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