A permease encoded by STL1 is required for active glycerol uptake by Candida albicans

Gerald Kayingo, António Martins, Rachael Andrie, Luisa Neves, Cândida Lucas, Brian Wong

Research output: Contribution to journalArticle

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Abstract

Candida albicans accumulates large amounts of the polyols glycerol and D-arabitol when the cells are exposed to physiological conditions relevant to stress and virulence in animals. Intracellular concentrations of glycerol are determined by rates of glycerol production and catabolism and of glycerol uptake and efflux through the plasma membrane. We and others have studied glycerol production in C. albicans, but glycerol uptake by C. albicans has not been studied. In the present study, we found that [14C]glycerol uptake by C. albicans SC5314 was (i) accumulative; (ii) dependent on proton-motive force; (iii) unaffected by carbon source; and (iv) unaffected by large molar excesses of D-arabitol or other polyols. The respective Km and Vmax values were 2.1 mM and 460 μmol h-1 (g dry wt)-1 in glucose medium and 2.6 mM and 268 μmol h-1 (g dry wt)-1 in glycerol medium. To identify the C. albicans glycerol uptake protein(s), we cloned the C. albicans homologues of the Saccharomyces cerevisiae genes GUP1 and STL1, both of which are known to be involved in glycerol transport. When multicopy plasmids encoding C. albicans STL1, C. albicans STL2 and C. albicans GUP1 were introduced into the corresponding S. cerevisiae null mutants, the transformants all acquired the ability to grow on minimal glycerol medium; however, only S. cerevisiae stl1 null mutants transformed with C. albicans STL1 actively took up extracellular [14C]glycerol. When both chromosomal alleles of C. albicans STL1 were deleted from C. albicans BWP17, the resulting stl1 null mutants grew poorly on minimal glycerol medium, and their ability to transport [14C]glycerol into the cell was markedly reduced. In contrast, deletion of both chromosomal alleles of C. albicans STL2 or of C. albicans GUP1 had no significant effects on [14C]glycerol uptake or the ability to grow on minimal glycerol medium. Northern blot analysis indicated that C. albicans STL1 was expressed in both glucose and glycerol media, conditions under which we detected wild-type active glycerol uptake. Furthermore, STL1 was highly expressed in salt-stressed cells; however, the stl1 null mutant was no more sensitive to salt stress than wild-type controls. We also detected high levels of STL2 expression in glycerol-grown cells, even though deletion of this gene did not influence glycerol uptake activity in glycerol-grown cells. We conclude from the results above that a plasma-membrane H+ symporter encoded by C. albicans STL1 actively transports glycerol into C. albicans cells.

Original languageEnglish (US)
Pages (from-to)1547-1557
Number of pages11
JournalMicrobiology
Volume155
Issue number5
DOIs
StatePublished - 2009

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Membrane Transport Proteins
Candida albicans
Glycerol
Saccharomyces cerevisiae
Salts
Alleles
Cell Membrane
Symporters

ASJC Scopus subject areas

  • Microbiology

Cite this

Kayingo, G., Martins, A., Andrie, R., Neves, L., Lucas, C., & Wong, B. (2009). A permease encoded by STL1 is required for active glycerol uptake by Candida albicans. Microbiology, 155(5), 1547-1557. https://doi.org/10.1099/mic.0.023457-0

A permease encoded by STL1 is required for active glycerol uptake by Candida albicans. / Kayingo, Gerald; Martins, António; Andrie, Rachael; Neves, Luisa; Lucas, Cândida; Wong, Brian.

In: Microbiology, Vol. 155, No. 5, 2009, p. 1547-1557.

Research output: Contribution to journalArticle

Kayingo, G, Martins, A, Andrie, R, Neves, L, Lucas, C & Wong, B 2009, 'A permease encoded by STL1 is required for active glycerol uptake by Candida albicans', Microbiology, vol. 155, no. 5, pp. 1547-1557. https://doi.org/10.1099/mic.0.023457-0
Kayingo, Gerald ; Martins, António ; Andrie, Rachael ; Neves, Luisa ; Lucas, Cândida ; Wong, Brian. / A permease encoded by STL1 is required for active glycerol uptake by Candida albicans. In: Microbiology. 2009 ; Vol. 155, No. 5. pp. 1547-1557.
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abstract = "Candida albicans accumulates large amounts of the polyols glycerol and D-arabitol when the cells are exposed to physiological conditions relevant to stress and virulence in animals. Intracellular concentrations of glycerol are determined by rates of glycerol production and catabolism and of glycerol uptake and efflux through the plasma membrane. We and others have studied glycerol production in C. albicans, but glycerol uptake by C. albicans has not been studied. In the present study, we found that [14C]glycerol uptake by C. albicans SC5314 was (i) accumulative; (ii) dependent on proton-motive force; (iii) unaffected by carbon source; and (iv) unaffected by large molar excesses of D-arabitol or other polyols. The respective Km and Vmax values were 2.1 mM and 460 μmol h-1 (g dry wt)-1 in glucose medium and 2.6 mM and 268 μmol h-1 (g dry wt)-1 in glycerol medium. To identify the C. albicans glycerol uptake protein(s), we cloned the C. albicans homologues of the Saccharomyces cerevisiae genes GUP1 and STL1, both of which are known to be involved in glycerol transport. When multicopy plasmids encoding C. albicans STL1, C. albicans STL2 and C. albicans GUP1 were introduced into the corresponding S. cerevisiae null mutants, the transformants all acquired the ability to grow on minimal glycerol medium; however, only S. cerevisiae stl1 null mutants transformed with C. albicans STL1 actively took up extracellular [14C]glycerol. When both chromosomal alleles of C. albicans STL1 were deleted from C. albicans BWP17, the resulting stl1 null mutants grew poorly on minimal glycerol medium, and their ability to transport [14C]glycerol into the cell was markedly reduced. In contrast, deletion of both chromosomal alleles of C. albicans STL2 or of C. albicans GUP1 had no significant effects on [14C]glycerol uptake or the ability to grow on minimal glycerol medium. Northern blot analysis indicated that C. albicans STL1 was expressed in both glucose and glycerol media, conditions under which we detected wild-type active glycerol uptake. Furthermore, STL1 was highly expressed in salt-stressed cells; however, the stl1 null mutant was no more sensitive to salt stress than wild-type controls. We also detected high levels of STL2 expression in glycerol-grown cells, even though deletion of this gene did not influence glycerol uptake activity in glycerol-grown cells. We conclude from the results above that a plasma-membrane H+ symporter encoded by C. albicans STL1 actively transports glycerol into C. albicans cells.",
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