1887

Microbiology Society logo

Volume 142, Issue 5

Demonstration of high-affinity Mn uptake in Saccharomyces cerevisiae: specificity and kinetics Free

Abstract

The existence of multiple transport systems for Mn in has been demonstrated in this study. Mn (supplied as MnCI) was accumulated by at all Mn concentrations examined (25 nM-1 mM) but a log-log plot of uptake rates and total amounts accumulated revealed the existence of at least two Mn concentration-dependent transport systems. Over a low Mn concentration range (25-1000 nM), high-affinity Mn uptake occurred with a value of 0.3 μM, while transformation of kinetic data obtained over the concentration range 5-200 μM revealed another system with a of 62 μM. Meaningful kinetic analyses were not possible at higher Mn concentrations because of toxicity: only about 30% of cells remained viable after 30 min incubation with 1000 μM MnCI. Release of K accompanied Mn accumulation and this increased with increasing Mn concentration. However, even in non-toxic Mn concentrations, the ratio of Mn uptake to K release greatly exceeded electroneutral stoichiometric exchange. In 50 μM MnCI, the ratio was 1: 123 and this increased to 1:2670 in 1000 μM MnCI, a toxic concentration. External Mg was found to decrease Mn accumulation at all concentrations examined, but to differing extents. Over the low Mn concentration range (5-200 μM), Mg competitively inhibited Mn uptake with a half-maximal inhibitory concentration, , of 5.5 μM Mg. However, even in the presence of a 50-fold excess of Mg, inhibition of Mn uptake was of the order of 72% and it appears that the cellular requirement for Mn could be maintained even in the presence of such a large excess of Mg. Over the high Mn concentration range (5-200 μM), the for Mg was 25.2 μM. At low Mn concentrations, Zn and Co, but not Cd, inhibited Mn uptake, which indicated that the high-affinity Mn uptake system was of low specificity, while at higher Mn concentrations, where the lower-affinity Mn transport system operated, inhibition was less marked. However, competition studies with potentially toxic metal cations were complicated due to toxic effects, particularly noticeable at 50 μM Co and Cd.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): divalent cations , Mg2+ , Mn2+ , Sattearomyces cerevisiae and transport kinetics
© Society for General Microbiology 1996
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-142-5-1159
1996-05-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/142/5/mic-142-5-1159.html?itemId=/content/journal/micro/10.1099/13500872-142-5-1159&mimeType=html&fmt=ahah

References

  1. Archibald F.S. Manganese: its acquisition by and function in the lactic acid bacteria. Crit Rev Microbiol 1986; 13:63–109
     [Google Scholar]
  2. Auling G. The effect of manganese limitation on DNA precursor biosynthesis during nucleotide fermentation with Brevi-bacterium ammoniagenes and Micrococcus luteus. Eur J Appi Microbiol Biotechnol 1983; 18:229–235
     [Google Scholar]
  3. Auling G. Manganese: function and transport in fungi. In Metal Ions in Fungi 1994 Edited by Winkelmann G., Winge D.R. New York: Marcel Dekker; pp 215–236
     [Google Scholar]
  4. Belde P.J.M., Kessels B.G.F., Moelans I.M., Borst-Pauwels G.W.F.H. Cd2+ uptake, Cd2+ binding and loss of cell K+ by a Cd-sensitive and a Cd-resistant strain of Saccharomyces cerevisiae. FEMS Microbiol Lett 1988; 49:493–498
     [Google Scholar]
  5. Bianchi M.E., Carbone M.L., Lucchini G. Mn2+ and Mg2+ uptake in Mn-sensitive and Mn-resistant yeast strains. Plant Sci Lett 1981a; 22:345–352
     [Google Scholar]
  6. Bianchi M.E., Carbone M.L., Lucchini G., Magni G.E. Mutants resistant to manganese in Saccharomyces cerevisiae. Curr Genet 1981b; 4:215–220
     [Google Scholar]
  7. Borst-Pauwels G.W.F.H. Ion transport in yeast. Biochim Biophys Acta 1981; 650:88–127
     [Google Scholar]
  8. Borst-Pauwels G.W.F.H. Simulation of all-or-none K+ efflux from yeast provoked by xenobiotics. Biochim Biophys Acta 1988; 937:88–92
     [Google Scholar]
  9. Borst-Pauwels G.W.F.H., Severens P.P.J. Effect of the surface potential upon ion selectivity found in competitive inhibition of divalent cation uptake. A theoretical approach. Physiol Plant 1984; 60:86–91
     [Google Scholar]
  10. Cornelius G., Nakashima H. Vacuoles play a decisive role in calcium homeostasis in Neurospora crassa. J Gen Microbiol 1987; 133:2341–2347
     [Google Scholar]
  11. De Rome L., Gadd G.M. Measurement of copper uptake in Saccharomyces cerevisiae using a Cu2+ selective electrode. FEMS Microbiol Lett 1987; 43:283–287
     [Google Scholar]
  12. Eilam Y., Lavi H., Grossowicz N. Cytoplasmic Ca2+ homeostasis maintained by a vacuolar Ca2+ transport system in the yeast Saccharomyces cerevisiae. J Gen Microbiol 1985; 131:623–629
     [Google Scholar]
  13. Failla M.L., Benedict C.D., Weinberg E.D. Accumulation and storage of Zn2+ by Candida utilis. J Gen Microbiol 1976; 94:23–26
     [Google Scholar]
  14. Frausto Da Silva J.J.R., Williams R.J.P. The Biological Chemistry of the Elements 1993 Oxford: Oxford University Press;
     [Google Scholar]
  15. Gadd G.M. The responses of fungi towards heavy metals. In Microbes in Extreme Environments 1986 Edited by Herbert R.A., Codd G.A. London: Academic Press; pp 83–110
     [Google Scholar]
  16. Gadd G.M. Interactions of fungi with toxic metals. New Phytol 1993; 124:25–60
     [Google Scholar]
  17. Gadd G.M., Mowll J.L. The relationship between cadmium uptake, potassium release and viability in Saccharomyces cerevisiae. FEMS Microbiol Lett 1983; 16:45–48
     [Google Scholar]
  18. Gadd G.M., White C. Heavy metal and radionuclide accumulation and toxicity in fungi and yeasts. In Metal-Microbe Interactions 1989 Edited by Poole R.K., Gadd G.M. Oxford: IRL Press; pp 19–38
     [Google Scholar]
  19. Garraway M.O., Evans R.C. Fungal Nutrition and Physiology 1984 New York: John Wiley;
     [Google Scholar]
  20. Hockertz S., Plonzig J., Auling G. Impairment of DNA formation is an early event in Aspergillus niger under manganese starvation. Appl Microbiol Biotechnol 1987a; 90:1420–1425
     [Google Scholar]
  21. Hockertz S., Schmid J., Auling G. A specific transport system for manganese in the filamentous fungus Aspergillus niger. J Gen Microbiol 1987b; 133:3513–3519
     [Google Scholar]
  22. Hughes M.N. The Inorganic Chemistry of Biological Processes 1990 Chichester: John Wiley;
     [Google Scholar]
  23. Jones R.P., Gadd G.M. Ionic nutrition of yeast - the physiological mechanisms involved and applications for biotechnology. Enzyme Microb Technol 1990; 12:402–418
     [Google Scholar]
  24. Kendrick M.J., May M.T., Plishka M.J., Robinson K.D. Metals in Biological Systems 1992 Chichester: Ellis Horwood;
     [Google Scholar]
  25. Kessels B.G.F., Belde P.J.M., Borst-Pauwels G.W.F.H. Protection of Saccharomyces cerevisiae against Cd2+ toxicity by Ca2+. J Gen Microbiol 1985; 131:2533–2537
     [Google Scholar]
  26. Kihn J.C., Dassargues C.M., Mestdagh M.M. Preliminary ESR study of Mn(II) retention by the yeast Saccharomyces. Can J Microbiol 1988; 34:1230–1234
     [Google Scholar]
  27. Kubicek C.P., Rohr M. Citric acid fermentation. Crit Rev Biotechnol 1986; 3:253–271
     [Google Scholar]
  28. Lawford H.G., Pik J.R., Lawford G.R., Williams T., Kligerman A. Hyperaccumulation of zinc by zinc-depleted Candida utilis grown in chemostat culture. Can J Microbiol 1980; 26:71–76
     [Google Scholar]
  29. Lichko L.P., Okorokov L.A., Kulaev I.S. Role of vacuolar ion pool in Saccharomyces carlsbergensis. Potassium efflux from vacuoles is coupled with manganese or magnesium influx. J Bacteriol 1980; 144:666–671
     [Google Scholar]
  30. Lin C.-M., Kosman D.J. Copper uptake in wild type and copper metallothionein-deficient Saccharomyces cerevisiae. J Biol Chem 1990; 265:9194–9200
     [Google Scholar]
  31. Meikle A.J., Reed R.H., Gadd G.M. Osmotic adjustment and the accumulation of organic solutes in whole cells and protoplasts of Saccharomyces cerevisiae. J Gen Microbiol 1988; 134:3049–3060
     [Google Scholar]
  32. Miller A.J., Vogg G., Sanders D. Cytosolic calcium homeostasis in fungi: roles of plasma membrane transport and intracellular sequestration of calcium. Proc Natl Acad Sei USA 1990; 87:9348–9352
     [Google Scholar]
  33. Nieuwenhuis B.J.W.M., Weijers A.G.M., Borst-Pauwels G.W.F.H. Uptake and accumulation of Mn2+ and Sr2+ in Saccharomyces cerevisiae. Biochim Biophys Acta 1981; 649:83–88
     [Google Scholar]
  34. Norris P.R., Kelly D.P. Accumulation of cadmium and cobalt by Saccharomyces cerevisiae. J Gen Microbiol 1977; 99:317–324
     [Google Scholar]
  35. Okorokov L.A. Main mechanisms of ion transport and regulation of ion concentrations in the yeast cytoplasm. In Environmental Regulation of Microbial Metabolism 1985 Edited by Kulaev I.S., Dawes E.A., Tempest D.W. London: Academic Press; pp 339–349
     [Google Scholar]
  36. Okorokov L.A., Lichko L.P., Kholodenko V.P., Kadomtseva V.M., Petrikevich S.B., Zaichkin E., Karimova A.M. Free and bound magnesium in fungi and yeasts. Folia Microbiol 1975; 20:460–466
     [Google Scholar]
  37. Okorokov L.A., Lichko L.P., Kulaev I.S. Vacuoles: main compartments of potassium, magnesium and phosphate ions in Saccharomyces carlsbergensis cells. J Bacteriol 1980; 144:661–665
     [Google Scholar]
  38. Okorokov L.A., Andreeva N.A., Lichko L.P., Valiakhmetov A.Y. Transmembrane gradient of K+ ions as an energy source in the yeast Saccharomyces carlsbergensis. Biochem Int 1983a; 6:463–472
     [Google Scholar]
  39. Okorokov L.A., Lichko L.P., Andreeva N.A. Changes of ATP, polyphosphate and K+ content in Saccharomyces cerevisiae during uptake of Mn2+ and glucose. Biochem Int 1983b; 6:481–488
     [Google Scholar]
  40. Okorokov L.A., Kulakovskaya T.V., Lichko L.P., Polorotova E.V. H+/ion antiport as the principal mechanism of transport systems in the vacuolar membrane of the yeast Saccharomyces carlsbergensis. FEBS Lett 1985; 192:303–306
     [Google Scholar]
  41. Parkin M.J., Ross I.S. Uptake of copper and manganese by the yeast, Candida utilis. Microbios Lett 1985; 29:115–120
     [Google Scholar]
  42. Parkin M.J., Ross I.S. The specific uptake of manganese in the yeast Candida utilis. J Gen Microbiol 1986; 132:2155–2160
     [Google Scholar]
  43. Perkins J., Gadd G.M. Caesium toxicity, accumulation and intracellular localization in yeasts. Mycol Res 1993; 97:717–724
     [Google Scholar]
  44. Pilz F., Auling G., Stephan D., Rau U., Wagner F. A high-affinity Zn2+ uptake system controls growth and biosynthesis of an extracellular, branched β-l, 3-/J-1, 6-glucan in Sclerotium rolfsii ATCC 15205. Exp Mycol 1991; 15:181–192
     [Google Scholar]
  45. Plonzig J., Auling G. Manganese deficiency impairs ribonucleotide reduction but not DNA replication in Arthrobacter species. Arch Microbiol 1987; 146:396–401
     [Google Scholar]
  46. Ramos S., Pena P., Valle E., Bergillos L., Parra F., Lazo P.S. Coupling of protons and potassium gradients in yeast. In Environmental Regulation of Microbial Metabolism 1985 Edited by Kulaev I.S., Dawes E.A., Tempest D.W. London: Academic Press; pp 351–357
     [Google Scholar]
  47. Ross I.S. Membrane transport processes and response to exposure to heavy metals. In Stress Tolerance of Fungi 1993 Edited by Jennings D.H. New York: Marcel Dekker; pp 97–125
     [Google Scholar]
  48. Sanders D. Kinetic modelling of plant and fungal membrane transport systems. Annu Rev Plant Physiol Plant Mol Biol 1990; 41:77–107
     [Google Scholar]
  49. Starling A.P., Ross I.S. Uptake of manganese by Penicillium notatum. Microbios 1990; 63:93–100
     [Google Scholar]
  50. White C., Gadd G.M. Uptake and cellular distribution of copper, cobalt and cadmium in strains of Saccharomyces cerevisiae cultured on elevated concentrations of these metals. FEMS Microbiol Ecol 1986; 38:277–283
     [Google Scholar]
  51. White C., Gadd G.M. The uptake and cellular distribution of zinc in Saccharomyces cerevisiae. J Gen Microbiol 1987a; 133:727–737
     [Google Scholar]
  52. White C., Gadd G.M. Inhibition of H+ efflux and K+ uptake and induction of K+ efflux in yeast by heavy metals. Toxic Assess 1987b; 2:437–444
     [Google Scholar]
  53. Williams R.J.P. Free manganese(II) and iron(II) cations can act as cellular controls. FEBS Lett 1982; 140:3–10
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-142-5-1159
Loading
Demonstration of high-affinity Mn2+ uptake in Saccharomyces cerevisiae: specificity and kinetics
Microbiology 142, 1159 (1996); https://doi.org/10.1099/13500872-142-5-1159
/content/journal/micro/10.1099/13500872-142-5-1159
/content/journal/micro/10.1099/13500872-142-5-1159
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error