Candida albicans produces and accumulates large amounts of the polyols d-arabitol and glycerol in culture, and/or in infected mammalian tissues. However, the effects of environmental stresses on production and accumulation of these polyols, and the means by which polyol production and accumulation are regulated have not been studied. C. albicans grown in glucose at 30 °C (i) produced maximal amounts of glycerol within 6 h, (ii) produced maximal amounts of d-arabitol and ribitol within 12 h, and (iii) released most of these polyols into the extracellular environment. C. albicans responded to osmotic and citric acid stress by producing and accumulating more glycerol, and to temperature and oxidative stresses by producing more d-arabitol. The increase in intracellular glycerol was proportional to extracellular osmolarity, suggesting that glycerol functions as an osmolyte. The MAP kinase Hog1p is required for wild-type glycerol production in several fungal species subjected to osmotic stress, but it is not known if Hog1p plays a role in regulating d-arabitol production. Therefore, two C. albicans hog1 null mutants were constructed and tested for the ability to produce glycerol and d-arabitol in response to environmental stresses. The ability to grow and produce glycerol when exposed to osmotic or citric acid stresses, and to produce d-arabitol when exposed to oxidative stress, was partially dependent on Hog1p, but the ability to produce d-arabitol when exposed to temperature stress was Hog1p independent. These results imply that multiple pathways regulate glycerol and d-arabitol synthesis in C. albicans.
AlbertynJ.,
HohmannS.,
TheveleinJ. M.,
PriorB. A.
1994; GPD1 , which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae , and its expression is regulated by the high-osmolarity glycerol response pathway. Mol Cell Biol 14:4135–4144
Alonso-MongeR.,
Navarro-GarciaF.,
MoleroG.,
Diez-OrejasR.,
GustinM.,
PlaJ.,
SanchezM.,
NombelaC.
1999; Role of the mitogen-activated protein kinase Hog1p in morphogenesis and virulence of Candida albicans
. J Bacteriol 181:3058–3068
Alonso-MongeR.,
NegredoA. I.,
EismanB.,
NombelaC.,
PlaJ, Navarro-GarcíaF.,
RomanE.2003; The Hog1 MAP kinase is essential in the oxidative stress response and chlamydospore formation in Candida albicans
. Eukaryot Cell 2:351–361[CrossRef]
AnsellR.,
GranathK.,
HohmannS.,
TheveleinJ. M.,
AdlerL.
1997; The two isoenzymes for yeast NAD+-dependent glycerol-3-phosphate dehydrogenase encoded by GPD1 and GPD2 have distinct roles in osmoadaption and redox regulation. EMBO J 16:2179–2187[CrossRef]
BernardE. M.,
ChristiansenL. J.,
TsangS. F.,
KiehnT. E.,
ArmstrongD.
1981; Rate of arabinitol production by pathogenic yeast species. J Clin Microbiol 14:189–194
BernardE. M.,
WongB.,
ArmstrongD.
1985; Stereoisomeric configuration of arabinitol in serum, urine, and tissues in invasive candidiasis. J Infect Dis 151:711–715[CrossRef]
BilslandE.,
MolinC.,
SwaminathanS.,
RamneA.,
SunnerhagenP.
2004; Rck1 and Rck2 MAPKAP kinases and the HOG pathway are required for oxidative stress resistance. Mol Microbiol 53:1743–1756[CrossRef]
ChaturvediV.,
WongB.,
NewmanS. L.
1996b; Oxidative killing of Cryptococcus neoformans by human neutrophils. Evidence that fungal mannitol protects by scavenging reactive oxygen intermediates. J Immunol 156:3836–3840
EdgleyM.,
BrownA. D.
1983; Yeast water relations: physiological changes induced by solute stress in Saccharomyces cerevisiae and Saccharomyces rouxii
. J Gen Microbiol 129:3453–3463
FanJ.,
WhitewayM.,
ShenS. H.
2005; Disruption of a gene encoding glycerol 3-phosphatase from Candida albicans impairs intracellular glycerol accumulation-mediated salt-tolerance. FEMS Microbiol Lett 245:107–116[CrossRef]
GillumA. M.,
TsayE. Y.,
KirschD. R.
1984; Isolation of the Candida albicans gene for orotidine-5′-phosphate decarboxylase by complementation of S. cerevisiae ura 3 and E. coli pyrF mutations. Mol Gen Genet 198:179–182[CrossRef]
HaghnazariE.,
HeyerW.-D.
2004; The Hog1 MAP kinase pathway and the Mec1 DNA damage checkpoint pathway independently control the cellular responses to hydrogen peroxide. DNA Repair 3:769–776[CrossRef]
JenningsD. B.,
EhrenshaftM.,
PharrD. M.,
WilliamsonJ. D.
1998; Roles for mannitol and mannitol dehydrogenases in active oxygen-mediated plant defense. Proc Natl Acad Sci U S A 95:15129–15133[CrossRef]
KayingoG.,
KilianS. G.,
PriorB. A.
2001; Conservation and release of osmolytes by yeasts during hypo-osmotic stress. Arch Microbiol 177:29–55[CrossRef]
KiehnT. E.,
BernardM.,
GoldJ. W. M.,
ArmstrongD.
1979; Candidiasis detection by gas-liquid chromatography of d-arabinitol, a fungal metabolite in human serum. Science 206:577–580[CrossRef]
LawrenceC. L.,
BottingC. H.,
AntrobusR.,
CooteP. J.
2004; Evidence of a new role for the high-osmolarity glycerol mitogen-activated protein kinase pathway in yeast: regulating adaptation to citric acid stress. Mol Cell Biol 24:3307–3323[CrossRef]
LeeL. K.,
BuckleyH. R.,
CampbellC. C.
1975; An amino acid liquid synthetic medium for the development of mycelial and yeast form of Candida albicans
. Sabouraudia 13:148–153[CrossRef]
NorbeckJ.,
BlombergA.,
AdlerL, PåhlmanA.-K.,
AkhtarN.1996; Purification and characterization of two isoenzymes of dl-glycerol-3-phosphatase from Saccharomyces cerevisiae . Identification of the corresponding GPP1 and GPP2 genes and evidence for osmotic regulation of Gpp2p expression by the osmosensing mitogen-activated protein kinase signal transduction pathway. J Biol Chem 271:13875–13881[CrossRef]
O'RourkeS. M.,
HerskowitzI.
2004; Unique and redundant roles for HOG MAPK pathway components as revealed by whole-genome expression analysis. Mol Biol Cell 15:532–532
PfallerM. A.,
JonesR. N.,
DoernG. V.,
SaderH. S.,
HollisR. J.,
MesserS. A.
for The SENTRY Participant Group1998; International surveillance of bloodstream infections due to Candida species: frequency of occurrence and antifungal susceptibilities of isolates collected in 1997 in the United States, Canada, and South America for the SENTRY Program. J Clin Microbiol 36:1886–1889
PosasF.,
ChamabersJ. R.,
HeymanJ. A.,
HoefflerJ. P.,
ArinoJ, de NadalE.2000; The transcriptional response of yeast to saline stress. J Biol Chem 275:17249–17255[CrossRef]
RepM.,
KrantzM.,
TheveleinJ. M.,
HohmannS.
2000; The transcriptional response of Saccharomyces cerevisiae to osmotic shock. Hot1p and Msn2p/Msn4p are required for the induction of subsets of high osmolarity glycerol pathway-dependent genes. J Biol Chem 275:8290–8300[CrossRef]
San JoséC.,
AlonsoR.,
Pérez-DíazR. M.,
PlaJ.,
NombelaC.
1996; The mitogen-activated protein kinase homolog HOG1 gene controls glycerol accumulation in the pathogenic fungus Candida albicans
. J Bacteriol 178:5850–5852
SmithD. A.,
NichollsS.,
MorganB. A.,
BrownA. J. P.,
QuinnJ.
2004; A conserved stress-activated protein kinase regulates a core stress response in the human pathogen Candida albicans
. Mol Biol Cell 15:4179–4190[CrossRef]
Van EckJ. H.,
PriorB. A.,
BrandtE. V.
1989; Accumulation of polyhydroxy alcohols by Hansenula anomala in response to water stress. J Gen Microbiol 135:1047–1054
WilsonR. B.,
DavisD.,
MitchellA. P.
1999; Rapid hypothesis testing in Candida albicans through gene disruption with short homology regions. J Bacteriol 181:1868–1874
WongB.,
BernardE. M.,
GoldJ. W.,
FongD.,
ArmstrongD.
1982; The arabinitol appearance rate in laboratory animals and humans: estimation from the arabinitol/creatinine ratio and relevance to the diagnosis of candidiasis. J Infect Dis 146:353–359[CrossRef]
WongB.,
MurrayJ. S.,
CastellanosM.,
CroenK. D.
1993; d-arabitol metabolism in Candida albicans : studies of the biosynthetic pathway and the gene that encodes NAD-dependent d-arabitol dehydrogenase. J Bacteriol 175:6314–6320
WongB.,
LeesonS.,
GrindleS.,
MageeB.,
BrooksE.,
MageeP. T.
1995; d-arabitol metabolism in Candida albicans : construction and analysis of mutants lacking d-arabitol dehydrogenase. J Bacteriol 177:2971–2976
YanceyP. H.,
ClarkM. E.,
HandS. C.,
BowlusR. D.,
SomeroG. N.
1982; Living with water stress: evolution of osmolyte systems. Science 217:1214–1222[CrossRef]