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

¹ Yale University;
² Departamento de Biologia/Universidade do Minho;
³ Division of Infectious Diseases, Department of Medicine, Oregon Health and Science University

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 studied glycerol production in Candida 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 2.2 mM and 460 µmol/h/g dry wt in glucose medium and 2.7 mM and 268 µmol/h/g dry wt in glycerol medium. To identify the C. albicans glycerol uptake protein(s), we cloned the C. albicans homologs of the Saccharomyces cerevisiae genes GUP1, STL1, and STL2. When multicopy plasmids encoding C. albicans GUP1, C. albicans STL1 and C. albicans STL2 were introduced into the corresponding S. cerevisiae null mutants, the transformants all acquired the ability to grow on minimal glycerol. 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 GUP1 or of C. albicans STL2 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 both in the presence of 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 mutant was no more sensitive to salt stress than wild-type controls. We also detected high levels of STL2 expression in glycerol, 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 proton symporter encoded by C. albicans STL1 actively transports glycerol into C. albicans cells.

⁴ E-mail: gerald.kayingo{at}yale.edu