Skip to main content

Functional Analysis of the Menkes Protein (MNK) Expressed from a cDNA Construct

  • Chapter
Book cover Copper Transport and Its Disorders

Abstract

A detailed structure/function analysis of the Menkes protein (MNK) is required to elucidate its role in maintaining cellular copper homeostasis. It was recently demonstrated that over-expression of the MNK protein confers a copper-resistance phenotype upon CHO-K1 cells (Camakaris et al, 1995), and that MNK in these cells is located primarily in the trans-Golgi network (Petris et al., 1996), but trafflcks to the plasma membrane in elevated copper levels. To investigate the molecular mechanism of the copper-induced trafficking of MNK, and copper translocation across cellular membranes, stable expression of the full length protein in mammalian cells is necessary, followed by the generation of cell lines that contain in vitro mutated MNK constructs.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Boshart, M., Weber, F., Jahn, G., Dorsch-Hasler, K., Fleckenstein, B., and Schaffner, W. (1985). A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell 41, 521–530.

    Article  PubMed  CAS  Google Scholar 

  • Brinster, R. L., Allen, J. ML, Behringer, R. R., Gelinas, R. E., and Palmiter, R. D. (1988). Introns increase transcriptional effciency in transgenic mice. Proc. Natl. Acad. Sci. USA 85, 836–840.

    Article  PubMed  CAS  Google Scholar 

  • Bull, P. C., and Cox, D. W. (1994). Wilson disease and Menkes disease: new handles on heavy-metal transport. Trends Genet. 10, 246–252.

    Article  PubMed  CAS  Google Scholar 

  • Camakaris, J., Petris, M. J., Bailey, L., Shen, P., Lockhart, P., Glover, T. W., Barcroft, C. L., Patton, J., and Mercer, J. F. B. (1995). Gene amplification of the Menkes (MNK; ATP7A) P-type ATPase gene of CHO cells is associated with copper resistance and enhanced copper efflux. Hum. Mol. Genet. 4, 2117–2123.

    Article  PubMed  CAS  Google Scholar 

  • Danks, D. M. (1995). Disorders of copper transport. In The Metabolic and Molecular Basis of Inherited Disease, C.R. Scriver, A.L. Beaudet, W.M. Sly and D. Valle, eds. (New York: McGraw-Hill), pp. 2211–2235.

    Google Scholar 

  • Das, S., Levinson, B., Vulpe, C., Whitney, S., Gitschier, J., and Packman, S. (1995). Similar splicing mutations of the Menkes/mottled copper-tranporting ATPase gene in occipital horn syndrome and the blotchy mouse. Am. J. Hum. Genet. 56, 570–579.

    PubMed  CAS  Google Scholar 

  • Dierick, H. A., Adam, A. N., Escara-Wilke, J. F., and Glover, T. W. (1997). Immunocytochemical localization of the Menkes copper transport protein (ATP7A) to the trans Golgi network. Hum. Mol. Genet. 6, 409–416.

    Article  PubMed  CAS  Google Scholar 

  • Francis, M. J., Jones, E., Levy, E. R., Chelly, J., and Monaco, A. P. (1996). Functional analysis of the Menkes disease gene. Am. J. Hum. Genet. 59 (suppl), A149.

    Google Scholar 

  • Gluzman, Y. (1981). SV40-transformed Simian cells support the replication of early SV40 mutants. Cell 23, 175–182.

    Article  PubMed  CAS  Google Scholar 

  • Gregory, R. G., Cheng, S. H., Rich, D. P., Marshall, J., Paul, S., Hehir, K., Ostegaard, L., Klinger, K. W., Welsh, M. J., and Smith, A. E. (1990). Expression and characterization of the cystic fibrosis transmembrane con-ductance regulator. Nature 347, 382–386.

    Article  PubMed  CAS  Google Scholar 

  • Griffiths, G., and Simons, K. (1986). The trans Golgi network: sorting at the exit site of of the Golgi complex. Science 234, 438–443.

    Article  PubMed  CAS  Google Scholar 

  • Herd, S. M., Camakaris, J., Christofferson, R., Wookey, P., and Danks, D. M. (1987). Uptake and efflux of copper-64 in Menkes’-disease and normal continuous lymphoid cell lines. Biochem. J. 247, 341–347.

    PubMed  CAS  Google Scholar 

  • Hung, I. H., Suzuki, M., Yamaguchi, Y., Yuan, D. S., Klausner, R. D., and Gitlin, J. D. (1997). Biochemical characterization of the Wilson disease protein and functional expression in the yeast Saccharomyces cerevisiae. J. Biol. Chem. 272, 21461–21466.

    Article  PubMed  CAS  Google Scholar 

  • Krauss, R. D., Bubien, J. K., Peiper, S. C., Collins, F. S., Kirk, K. L., Frizzell, R. A., and Rado, T. A. (1992). Transfection of wild-type CFTR into cystic fibrosis lymphocytes restores chloride conductance at G1 of the cell cycle. Embo J. 11, 875–883.

    PubMed  CAS  Google Scholar 

  • Kriegler, M. P. (1990). Gene transfer and expression. A laboratory manual. (New York: Freeman).

    Google Scholar 

  • Kuivaniemi, H., Peltonen, L., and Kivirikko, K. I. (1985). Type IX Ehlers-Danlos syndrome and Menkes syndrome: the decrease in lysyl oxidase activity is associated with a corresponding deficiency in the enzyme protein. Am. J. Hum. Genet. 37, 798–808.

    PubMed  CAS  Google Scholar 

  • La Fontaine, S., Firth, S. D., Lockhart, P. J., Brooks, H., Parton, R. G., Camakaris, J., and Mercer, J. F. B. (1997). Functional analysis and intracellular localization of the human Menkes Disease protein (MNK) stably expressed from a cDNA construct in Chinese Hamster Ovary cells (CHO-K1). Submitted for publication.

    Google Scholar 

  • La Fontaine, S., Firth, S. D., Lockhart, P. J., Paynter, J. A., and Mercer, J. F. B. (1997). Low copy number plasmid vectors for eukaryotic gene expression: transient expression of the Menkes protein. Plasmid. (Accepted for publication).

    Google Scholar 

  • Lippincott-Schwartz, J., Yuan, L., Tipper, C., Amherdt, M., Orci, L., and Klausner, R. D. (1991). Brefeldin A’s effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffick. Cell 67, 601–616.

    Article  PubMed  CAS  Google Scholar 

  • Lutsenko, S., Petrukhin, K., Cooper, M. J., Gilliam, C. T, and Kaplan, J. H. (1997). N-terminal domains of human copper-transporting adenosine triphosphatases (the Wilson’s and Menkes disease proteins) bind copper selectively in vivo and in vitro with stoichiometry of one copper per metal-binding repeat. J. Biol. Chem. 272, 18939–18944.

    Article  PubMed  CAS  Google Scholar 

  • Maruyama, K., and MacLennan, D. H. (1988). Mutation of aspartic acid-351, lysine-352, and lysine-515 alters the Ca2+ transport activity of the Ca2+-ATPase expressed in COS-1 cells. Proc. Natl. Acad. Sci. USA 85, 3314–3318.

    Article  PubMed  CAS  Google Scholar 

  • Morelle, G. (1989). A plasmid extraction procedure on a miniprep scale. Focus 11, 7–8.

    Google Scholar 

  • Pelham, H. R. B. (1991). Multiple targets for Brefeldin A. Cell 67, 449–451.

    Article  PubMed  CAS  Google Scholar 

  • Peterson, M. G., Hannan, F., and Mercer, J. F. B. (1988). The sheep metallothionein gene family. Eur. J. Biochem. 774, 417–424.

    Article  Google Scholar 

  • Petris, M. J., Mercer, J. F. B., Culvenor, J. G., Lockhart, P., Gleeson, P. A., and Camakaris, J. (1996). 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. 15, 6084–6095.

    PubMed  CAS  Google Scholar 

  • Reaves, B., and Banting, G. (1992). Perturbation of the morphology of the trans-Golgi network following Brefeldin A treatment: redistribution of a TGN-specific integral membrane protein, TGN38. J. Cell Biol. 116, 85–94.

    Article  PubMed  CAS  Google Scholar 

  • Royce, P. M., Camakaris, J., Mann, J. R., and Danks, D. M. (1982). Copper metabolism in mottled mouse mutants. Biochem. J. 202, 369–371.

    PubMed  CAS  Google Scholar 

  • Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989). Molecular cloning. A laboratory manual., 2nd Edition (New York: Cold Spring Harbor Laboratory Press).

    Google Scholar 

  • Solioz, M., and Vulpe, C. (1996). CPX-type ATPases: a class of P-type ATPases that pump heavy metals. Trends Biochem. Sci. 21, 237–241.

    PubMed  CAS  Google Scholar 

  • Stearman, R., Yuan, D. S., Yamaguchi-Iwai, Y, Klausner, R. D., and Dancis, A. (1996). A permease-oxidase complex involved in high-affinity iron uptake in yeast. Science 271, 1552–1557.

    Article  PubMed  CAS  Google Scholar 

  • Thomsen, D. R., Stenberg, R. M., Goins, W. F., and Stinski, M. F. (1984). Promoter-regulatory region of the major immediate early gene of human cytomegalovirus. Proc. Natl. Acad. Sci. USA 81, 659–663.

    Article  PubMed  CAS  Google Scholar 

  • Versieck, J., and Cornelis, R. (1980). Normal levels of trace elements in human blood plasma or serum. Anal. Chim., Acta 116, 217–254.

    Article  CAS  Google Scholar 

  • Vieira, J., and Messing, J. (1982). The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19, 259–268.

    Article  PubMed  CAS  Google Scholar 

  • Vulpe, C. D., and Packman, S. (1995). Cellular copper transport. Annu. Rev. Nutr. 15, 293–322.

    Article  PubMed  CAS  Google Scholar 

  • Walls, J. D., Berg, D. T., Yan, S. B., and Grinnell, B. W. (1989). Amplification of multicistronic plasmids in the human 293 cell line and secretion of correctly processed recombinant human protein C. Gene 81, 139–149.

    Article  PubMed  CAS  Google Scholar 

  • Wang, R. F., and Kushner, S. R. (1991). Construction of versatile low-copy-number vectors for cloning, sequencing and gene expression in Escherichia coli. Gene 100, 195–199.

    Article  PubMed  CAS  Google Scholar 

  • Yamaguchi, Y., Heiny, M. E., Suzuki, M., and Gitlin, J. D. (1997). Biochemical characterization and intracellular localization of the Menkes disease protein. Proc. Natl. Acad. Sci. USA 93, 14030–14035.

    Article  Google Scholar 

  • Yuan, D. S., Dancis, A., and Klausner, R. D. (1997). Restriction of copper export in Saccharomyces cerevisiae to a late Golgi or post-Golgi compartment in the secretory pathway. J. Biol. Chem. 272, 25787–25793.

    Article  PubMed  CAS  Google Scholar 

  • Yuan, D. S., Stearman, R., Dancis, A., Dunn, T., Beeler, T., and Klausner, R. D. (1995). The Menkes/Wilson disease gene homologue in yeast provides copper to a ceruloplasmin-like oxidase required for iron uptake. Proc. Natl. Acad. Sci. USA 92, 2632–2636.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1999 Springer Science+Business Media New York

About this chapter

Cite this chapter

La Fontaine, S., Firth, S.D., Lockhart, P.J., Brooks, H., Camakaris, J., Mercer, J.F.B. (1999). Functional Analysis of the Menkes Protein (MNK) Expressed from a cDNA Construct. In: Leone, A., Mercer, J.F.B. (eds) Copper Transport and Its Disorders. Advances in Experimental Medicine and Biology, vol 448. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-4859-1_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-4615-4859-1_6

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4613-7204-2

  • Online ISBN: 978-1-4615-4859-1

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics