1887

Abstract

The basis of fructophily in the yeast has been shown to reside in the performance of transport systems for hexoses. In this study, a gene encoding a fructose-specific transporter was characterized. The strategy involved the functional complementation of a strain that does not take up hexoses ( strain). This strain was transformed with a genomic library of . One transformant capable of growing on fructose, but not on glucose, was obtained. This transformant did not transport -[C]glucose, and the kinetic parameters for -[C]fructose were =3·3 mmol h g and =80·4 mM. As in the original strain of , fructose uptake was not inhibited by the presence of other hexoses or uranyl. The plasmid responsible for the observed phenotype was found to carry an ORF encoding a 616 amino acid protein with the characteristics of a membrane transporter, which was designated (fructose facilitator ). The impairment in function observed in an transformant expressing a truncated Ffz1 protein lacking 67 amino acids at the C-terminus suggests an important role for this terminal part in the proper structure of the transporter.

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2004-07-01
2020-08-12
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References

  1. Agatep R., Kirkpatric R. D., Prchaliuk D. L., Woods R. A., Gietz R. D.. 1998; Transformation of Saccharomyces cerevisiae by lithium acetate/single-stranded carrier DNA/polyethylene glycol (LiAc/ss-DNA/PEG) protocol. Technical Tips Onlinehttp://www.ciwemb.edu/labs/koshland/Protocols/YEAST/LiAc.html
    [Google Scholar]
  2. Boeke J. D., Trueheart J., Natsoulis G., Fink G. R.. 1987; 5-Fluoroorotic acid as a selective agent in yeast molecular genetics. Methods Enzymol154:164–175
    [Google Scholar]
  3. Cason D. T., Spencer-Martins I., van Uden N.. 1986; Transport of fructose by a proton symport in a brewing yeast. FEMS Microbiol Lett36:307–309[CrossRef]
    [Google Scholar]
  4. Cirillo V. P.. 1968; Relationship between sugar structure and competition for the sugar transport system in baker's yeast. J Bacteriol95:603–611
    [Google Scholar]
  5. Diezemann A., Boles E.. 2003; Functional characterization of the Frt1 sugar transporter and of fructose uptake in Kluyveromyces lactis. Curr Genet43:281–288[CrossRef]
    [Google Scholar]
  6. Fan J., Charturvedi V., Shen S. H.. 2002; Identification and phylogenetic analysis of a glucose transporter gene family from the human pathogenic yeast Candida albicans. J Mol Evol55:336–346[CrossRef]
    [Google Scholar]
  7. Fleet G.. 1992; Spoilage yeasts. Crit Rev Biotechnol12:1–44[CrossRef]
    [Google Scholar]
  8. Fuhrmann G. F., Storch D., Bode H.-P., Völker B.. 1992; Inhibition of glucose transport in Saccharomyces cerevisiae by uranyl ions. J Biotechnol27:75–84[CrossRef]
    [Google Scholar]
  9. Gonçalves P., Rodrigues de Sousa H., Spencer-Martins I.. 2000; FSY1, a novel gene encoding a specific fructose/H+ symporter in the type strain of Saccharomyces carlsbergensis. J Bacteriol182:5628–5630[CrossRef]
    [Google Scholar]
  10. Hoffman C. S., Winston F.. 1987; A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene57:267–272[CrossRef]
    [Google Scholar]
  11. Kotyk A.. 1967; Properties of the sugar carrier in baker's yeast. Folia Microbiol12:121–131[CrossRef]
    [Google Scholar]
  12. Loureiro-Dias M. C.. 1987; Glucose and polyol transport systems in Candida intermedia and their regulation. J Gen Microbiol133:2737–2742
    [Google Scholar]
  13. Peinado J. M., Cameira-dos-Santos P. J., Loureiro-Dias M. C.. 1989; Regulation of glucose transport in Candida utilis. J Gen Microbiol135:195–201
    [Google Scholar]
  14. Reifenberger E., Boles E., Ciriacy M.. 1997; Kinetic characterization of individual hexose transporters of Saccharomyces cerevisiae and their relation to the triggering mechanisms of glucose repression. Eur J Biochem245:324–333[CrossRef]
    [Google Scholar]
  15. Rodrigues F., Zeeman A. M., Alves C., Sousa M. J., Steensman H. Y., Côrte-Real M., Leão C.. 2001; Construction of a genomic library of the food spoilage yeast Zygosaccharomyces bailii and isolation of the β-isopropylamate dehydrogenase gene (ZbLEU2. FEMS Yeast Res1:67–71
    [Google Scholar]
  16. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
  17. Sikorski R. S., Hieter P.. 1989; A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics122:19–27
    [Google Scholar]
  18. Sousa-Dias S., Gonçalves T., Leyva J. S., Peinado J. M., Loureiro-Dias M. C.. 1996; Kinetics and regulation of fructose and glucose transport systems are responsible for fructophily in Zygosaccharomyces bailii. Microbiology142:1733–1738[CrossRef]
    [Google Scholar]
  19. Spencer-Martins I., van Uden N.. 1985; Catabolite interconversion of glucose transport systems in the yeast Candida wickerhamii. Biochim Biophys Acta812:168–172[CrossRef]
    [Google Scholar]
  20. Wieczorke R., Krampe S., Weierstall T., Freidel K., Hollenberg C. P., Boles E.. 1999; Concurrent knock-out of at least 20 transporter genes is required to block uptake of hexoses in Saccharomyces cerevisiae. FEBS Lett464:123–128[CrossRef]
    [Google Scholar]
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