Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Manipulation of intracellular magnesium levels in Saccharomyces cerevisiae with deletion of magnesium transporters

Abstract

Magnesium is an important divalent ion for organisms. There have been a number of studies in vitro suggesting that magnesium affects enzyme activity. Surprisingly, there have been few studies to determine the cellular mechanism for magnesium regulation. We wished to determine if magnesium levels could be regulated in vivo. It is known that Saccharomyces cerevisiae has two magnesium transporters (ALR1 and ALR2) across the plasma membrane. We created S. cerevisiae strains with deletion of one (alr1 or alr2) or both (alr1 alr2) transporters. The deletion of ALR1 resulted in a decrease in intracellular magnesium levels. An increase from 5 to 100 mM in the exogenous magnesium level increased the intracellular levels of magnesium in the alr1 and alr1 alr2 strains, whereas the expression of magnesium transporters from S. cerevisiae or Arabidopsis thaliana led to a change of the intracellular levels of magnesium in those strains. The deletion of magnesium transporters in A. cerevisiae and overexpression of magnesium transporters from A. thaliana also affected the intracellular concentrations of a range of metal ions, which suggests that cells use non-specific transporters to help regulate metal homeostasis.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Notes

  1. 1.

    The abbreviations used here are CSM, complete supplement media; PCR, polymerase chain reaction; bp, base pair; and kb, kilobase.

References

  1. Brandt DR, Ross EM (1986) Catecholamine-stimulated GTPase cycle. Multiple sites of regulation by beta-adrenergic receptor and Mg2+ studied in reconstituted receptor-Gs vesicles. J Biol Chem 261:1656–1664

  2. Bui DM et al (1999) The bacterial magnesium transporter CorA can functionally substitute for its putative homologue Mrs2p in the yeast inner mitochondrial membrane. J Biol Chem 274:20438–20443

  3. Butt TR et al (1984) Copper metallothionein of yeast, structure of the gene, and regulation of expression. A Proc Natl Acad Sci USA 81:3332–3336

  4. Cech SY, Maguire ME (1982) Magnesium regulation of the beta-receptor-adenylate cyclase complex .1. Effects of manganese on receptor-binding and cyclase activation. Mol Pharmacol 22:267–273

  5. Cech SY et al (1980) Adenylate-cyclase—the role of magnesium and other divalent-cations. Mol Cell Biochem 33:67–92

  6. Chamnongpol S, Groisman EA (2002) Mg2+ homeostasis and avoidance of metal toxicity. Mol Microbiol 44:561–571

  7. Culotta VC et al (1994) CRS5 encodes a metallothionein-like protein in Saccharomyces cerevisiae. J Biol Chem 269:25295–25302

  8. Culotta VC et al (2005) Manganese transport and trafficking: lessons learned from Saccharomyces cerevisiae. Eukaryot Cell 4:1159–1165

  9. da Costa BM et al (2005) Regulation of rubber biosynthetic rate and molecular weight in Hevea brasiliensis by metal cofactor. Biomacromolecules 6:279–289

  10. da Costa BMT et al (2006) Magnesium ion regulation of in vitro rubber biosynthesis by Parthenium argentatum Gray. Phytochemistry 67:1621–1628

  11. Dancis A et al (1994) S. Molecular characterization of a copper transport protein in S. cerevisiae: an unexpected role for copper in iron transport. Cell 76:393–402

  12. Dennis PB et al (2001) T.O.R. Mammalian, a homeostatic ATP sensor. Science 294:1102–1105

  13. Drummond RSM et al (2006) A putative magnesium transporter AtMRS2-11 is localized to the plant chloroplast envelope membrane system. Plant Sci 170:78–89

  14. Eide DJ (1998) The molecular biology of metal ion transport in Saccharomyces cerevisiae. Annu Rev Nutr 18:441–469

  15. Eide DJ et al (2005) Characterization of the yeast ionome: a genome-wide analysis of nutrient mineral and trace element homeostasis in Saccharomyces cerevisiae. Genome Biol 6:R77

  16. Gardner RC (2003) Genes for magnesium transport. Curr Opin Plant Biol 6:263–267

  17. Gietz RD, Woods RA (2002) Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Guide to Yeast Genetics and Molecular and Cell Biology B Pt 350:87–96

  18. Graschopf A et al (2001) The yeast plasma membrane protein Alr1 controls Mg2+ homeostasis and is subject to Mg2+-dependent control of its synthesis and degradation. J Biol Chem 276:16216–16222

  19. Gregan J et al (2001) The mitochondrial inner membrane protein Lpe10p, a homologue of Mrs2p, is essential for magnesium homeostasis and group II intron splicing in yeast. Mol Gen Genet 264:773–781

  20. Higashijima T et al (1987) Effects of Mg2+ and the beta gamma-subunit complex on the interactions of guanine nucleotides with G proteins. J Biol Chem 262:762–766

  21. Horlitz M, Klaff P (2000) Gene-specific trans-regulatory functions of magnesium for chloroplast mRNA stability in higher plants. J Biol Chem 275:35638–35645

  22. Karin M et al (1984) Primary structure and transcription of an amplified genetic locus: the CUP1 locus of yeast. A Proc Natl Acad Sci USA 81:337–341

  23. Li L, Kaplan J (1998) Defects in the yeast high affinity iron transport system result in increased metal sensitivity because of the increased expression of transporters with a broad transition metal specificity. J Biol Chem 273:22181–22187

  24. Li LG et al (2001) A novel family of magnesium transport genes in arabidopsis. Plant Cell 13:2761–2775

  25. Liu XF, Culotta VC (1999a) Mutational analysis of Saccharomyces cerevisiae Smf1p, a member of the Nramp family of metal transporters. J Mol Biol 289:885–891

  26. Liu XF, Culotta VC (1999b) Post-translation control of Nramp metal transport in yeast. Role of metal ions and the BSD2 gene. J Biol Chem 274:4863–4868

  27. Liu GJ et al (2002) Large Mg2+-dependent currents are associated with the increased expression of ALR1 in Saccharomyces cerevisiae. FEMS Microbiol Lett 213:231–237

  28. MacDiarmid CW, Gardner RC (1998) Overexpression of the Saccharomyces cerevisiae magnesium transport system confers resistance to aluminum ion. J Biol Chem 273:1727–1732

  29. MacDiarmid CW et al (2000) Zinc transporters that regulate vacuolar zinc storage in Saccharomyces cerevisiae. EMBO J 19:2845–2855

  30. Maguire ME (1984) Hormone-sensitive magnesium transport and magnesium regulation of adenylate-cyclase. Trends Pharmacol Sci 5:73–77

  31. Meijer AH et al (1998) Vectors for transcription factor cloning and target site identification by means of genetic selection in yeast. Yeast 14:1407–1415

  32. Moncrief MBC, Maguire ME (1999) Magnesium transport in prokaryotes. J Biol Inorg Chem 4:523–527

  33. Montell C (2003) Mg2+ homeostasis: the Mg(2+)nificent TRPM chanzymes. Curr Biol 13:R799–R801

  34. Nadler MJS et al (2001) LTRPC7 is a Mg center dot ATP-regulated divalent cation channel required for cell viability. Nature 411:590–595

  35. Portnoy ME et al (2000) Saccharomyces cerevisiae expresses three functionally distinct homologues of the nramp family of metal transporters. Mol Cell Biol 20:7893–7902

  36. Radisky D, Kaplan J (1999) Regulation of transition metal transport across the yeast plasma membrane. J Biol Chem 274:4481–4484

  37. Romani AMP, Maguire ME (2002) Hormonal regulation of Mg2+ transport and homeostasis in eukaryotic cells. Biometals 15:271–283

  38. Rubin H (1975) Central role for magnesium in coordinate control of metabolism and growth in animal cells. Proc Natl Acad Sci USA 72:3551–3555

  39. Rubin H (2005) Magnesium: the missing element in molecular views of cell proliferation control. Bioessays 27:311–320

  40. Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor

  41. Sanui H, Rubin AH (1978) Membrane bound and cellular cationic changes associated with insulin stimulation of cultured cells. J Cell Physiol 96:265–278

  42. Schock I et al (2000) A member of a novel Arabidopsis thaliana gene family of candidate Mg2+ ion transporters complements a yeast mitochondrial group II intron-splicing mutant. Plant J 24:489–501

  43. Shaul O (2002) Magnesium transport and function in plants: the tip of the iceberg. Biometals 15:309–323

  44. Stearman R et al (1998) YIpDCE1—an integrating plasmid for dual constitutive expression in yeast. Gene 212:197–202

  45. Wachek M et al (2006) Oligomerization of the Mg2+-transport proteins Alr1p and Alr2p in yeast plasma membrane. FEBS J 273:4236–4249

  46. Walker GM, Duffus JH (1980) Magnesium-ions and the control of the cell-cycle in yeast. J Cell Sci 42:329–356

  47. Zhao H, Eide D (1996) The yeast ZRT1 gene encodes the zinc transporter protein of a high-affinity uptake system induced by zinc limitation. Proc Natl Acad Sci USA 93:2454–2458

  48. Zsurka G et al (2001) The human mitochondrial Mrs2 protein functionally substitutes for its yeast homologue, a candidate magnesium transporter. Genomics 72:158–168

Download references

Acknowledgment

We thank Prof. Luan, Department of Plant and Microbial Biology, University of California, Berkeley, and Prof. Weis, Department of Molecular and Cell Biology, University of California, Berkeley, for kindly providing plasmid pMGT10 and strain BY4741, respectively.

Author information

Correspondence to Jay D. Keasling.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplemental Material

(DOC 241 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

da Costa, B.M.T., Cornish, K. & Keasling, J.D. Manipulation of intracellular magnesium levels in Saccharomyces cerevisiae with deletion of magnesium transporters. Appl Microbiol Biotechnol 77, 411–425 (2007). https://doi.org/10.1007/s00253-007-1177-4

Download citation

Keywords

  • Magnesium
  • Saccharomyces cerevisiae
  • alr1
  • alr2