The study of gene regulation in many organisms has been facilitated by the development of reporter genes. The authors report the use of lacZ from Streptococcus thermophilus, a gene encoding a β-galactosidase, as a reporter for the fungal pathogen Candida albicans. As test cases, Strep. thermophilus lacZ was placed under control of three different C. albicans promoters: MAL2 (maltase), inducible by maltose; HWP1 (hyphal cell wall protein), induced by conditions that promote filamentous growth; and ACT1 (actin). These constructs were each integrated into the C. albicans genome and β-galactosidase activity was readily detected from these strains, but only under the appropriate growth conditions. β-Galactosidase activity could be detected by several methods: quantitative liquid assays using permeabilized cells, colorimetric assays of colonies replicated to paper filters, and in situ coloration of colonies growing on medium containing the indicator X-Gal. These results show the usefulness of Strep. thermophilus lacZ as a monitor of gene regulation in this medically important yeast.
AlexL. A.,
KorchC.,
SelitrennikoffC. P.,
SimonM. I.
1998; COS1, a two-component histidine kinase that is involved in hyphal development in the opportunistic pathogen Candida albicans. Proc Natl Acad Sci USA 95:7069–7073[CrossRef]
AusubelF. M.,
BrentR.,
KingstonR. E.,
MooreD. D.,
SeidmanJ. G.,
SmithJ. A.,
StruhlK.
1992Current Protocols in Molecular Biology New York: Greene Publishing Associates and Wiley-Interscience;
BlancoC.,
RitzenthalerP.,
Mata-GilsingerM.
1985; Nucleotide sequence of a regulatory region of the uidA gene in Escherichia coli K12. Mol Gen Genet 199:101–105[CrossRef]
BrownD. H.Jr,
SlobodkinI. V.,
KumamotoC. A.
1996; Stable transformation and regulated expression of an inducible reporter construct in Candida albicans using restriction enzyme-mediated integration. Mol Gen Genet 251:75–80
CaleraJ. A.,
ZhaoX. J.,
CalderoneR.
2000; Defective hyphal development and avirulence caused by a deletion of the SSK1 response regulator gene in Candida albicans. Infect Immun 68:518–525[CrossRef]
CormackB. P.,
BertramG.,
EgertonM.,
GowN. A. R.,
FalkowS.,
BrownA. J.
1997; Yeast-enhanced green fluorescent protein (yEGFP): a reporter of gene expression in Candida albicans. Microbiology 143:303–311[CrossRef]
CsankC.,
MakrisC.,
MelocheS.,
DignardD.,
ThomasD. Y.,
WhitewayM,
SchröppelK.,
RöllinghoffM.1997; Derepressed hyphal growth and reduced virulence in a VH1 family-related protein phosphatase mutant of the human pathogen Candida albicans. Mol Biol Cell. 82539–2551[CrossRef]
CsankC.,
SchroppelK.,
LebererE.,
HarcusD.,
MohamedO.,
MelocheS.,
ThomasD. Y.,
WhitewayM.
1998; Roles of the Candida albicans mitogen-activated protein kinase homolog, Cek1p, in hyphal development and systemic candidiasis. Infect Immun 66:2713–2721
DelbruckS.,
ErnstJ. F.
1993; Morphogenesis-independent regulation of actin transcript levels in the pathogenic yeast Candida albicans. Mol Microbiol. 10859–866[CrossRef]
FonziW. A.,
IrwinM. Y.
1993; Isogenic strain construction and gene mapping in Candida albicans. Genetics 134:717–728
GaleC. A.,
BendelC. M.,
McClellanM.,
HauserM.,
BeckerJ. M.,
BermanJ.,
HostetterM. K.
1998; Linkage of adhesion, filamentous growth, and virulence in Candida albicans to a single gene, INT1. Science 279:1355–1358[CrossRef]
GietzR. D.,
SchiestlR. H.,
WillemsA. R.,
WoodsR. A.
1995; Studies on the transformation of intact yeast cells by the LiAc/SS-DNA/PEG procedure. Yeast 11:355–360[CrossRef]
GimenoC. J.,
LjungdahlP. O.,
StylesC. A.,
FinkG. R.
1992; Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: regulation by starvation and RAS . Cell 68:1077–1090[CrossRef]
GuarenteL.,
PtashneM.
1981; Fusion of Escherichia coli lacZ to the cytochrome c gene of Saccharomyces cerevisiae. Proc Natl Acad Sci USA 78:2199–2203[CrossRef]
HillJ.,
DonaldK. A.,
GriffithsD. E.,
DonaldG.
1991; DMSO-enhanced whole cell yeast transformation. [An erratum appears in Nucleic Acids Res 11, 6688.]. Nucleic Acids Res 19:5791[CrossRef]
JacobsonR. H.,
ZhangX. J.,
DuBoseR. F.,
MatthewsB. W.
1994; Three-dimensional structure of beta-galactosidase from E. coli. Nature 369:761–766[CrossRef]
LebererE.,
HarcusD.,
BroadbentI. D.7 other authors1996; Signal transduction through homologues of the Ste20p and Ste7p protein kinases can trigger hyphal formation in the pathogenic fungus Candida albicans. Proc Natl Acad Sci USA 93:13217–13222[CrossRef]
LeukerC. E.,
HahnA. M.,
ErnstJ. F.
1992; β-Galactosidase of Kluyveromyces lactis (Lac4p) as reporter of gene expression in Candida albicans and C. tropicalis. Mol Gen Genet 235:235–241[CrossRef]
LeukerC. E.,
SonnebornA.,
DelbruckS.,
ErnstJ. F.
1997; Sequence and promoter regulation of the PCK1 gene encoding phosphoenolpyruvate carboxykinase of the fungal pathogen Candida albicans. Gene 192:235–240[CrossRef]
LoH. J.,
KohlerJ. R.,
DiDomenicoB.,
LoebenbergD.,
CacciapuotiA.,
FinkG. R.
1997; Nonfilamentous C. albicans mutants are avirulent. Cell 90:939–949[CrossRef]
OhamaT.,
SuzukiT.,
MoriM.,
OsawaS.,
UedaT.,
WatanabeK.,
NakaseT.
1993; Non-universal decoding of the leucine codon CUG in several Candida species. Nucleic Acids Res 21:4039–4045[CrossRef]
RoseM.,
CasadabanM. J.,
BotsteinD.
1981; Yeast genes fused to beta-galactosidase in Escherichia coli can be expressed normally in yeast. Proc Natl Acad Sci USA 78:2460–2464[CrossRef]
SchroederC. J.,
RobertC.,
LenzenG.,
McKayL. L.,
MercenierA.
1991; Analysis of the lacZ sequences from two Streptococcus thermophilus strains: comparison with the Escherichia coli and Lactobacillus bulgaricus β-galactosidase sequences. J Gen Microbiol 137:369–380[CrossRef]
SreekrishnaK.,
DicksonR. C.
1985; Construction of strains of Saccharomyces cerevisiae that grow on lactose. Proc Natl Acad Sci USA 82:7909–7913[CrossRef]
SrikanthaT.,
KlapachA.,
LorenzW. W.,
TsaiL. K.,
LaughlinL. A.,
GormanJ. A.,
SollD. R.
1996; The sea pansy Renilla reniformis luciferase serves as a sensitive bioluminescent reporter for differential gene expression in Candida albicans. J Bacteriol 178:121–129
SrikanthaT.,
TsaiL. K.,
SollD. R.
1997; The WHI1 gene of Candida albicans is regulated in two distinct developmental programs through the same transcription activation sequences. J Bacteriol 179:3837–3844
StaabJ. F.,
FerrerC. A.,
SundstromP.
1996; Developmental expression of a tandemly repeated, proline- and glutamine-rich amino acid motif on hyphal surfaces of Candida albicans. J Biol Chem 271:6298–6305[CrossRef]
StoldtV. R.,
SonnebornA.,
LeukerC. E.,
ErnstJ. F.
1997; Efg1p, an essential regulator of morphogenesis of the human pathogen Candida albicans , is a member of a conserved class of bHLH proteins regulating morphogenetic processes in fungi. EMBO J 16:1982–1991[CrossRef]
SwobodaR. K.,
BertramG.,
DelbruckS.,
ErnstJ. F.,
GowN. A. R.,
GoodayG. W.,
BrownA. J.
1994; Fluctuations in glycolytic mRNA levels during morphogenesis in Candida albicans reflect underlying changes in growth and are not a response to cellular dimorphism. Mol Microbiol 13:663–672[CrossRef]
TimpelC.,
Strahl-BolsingerS.,
ZiegelbauerK.,
ErnstJ. F.
1998; Multiple functions of Pmt1p-mediated protein O-mannosylation in the fungal pathogen Candida albicans. J Biol Chem 273:20837–20846[CrossRef]
WirschingS.,
MichelS.,
KohlerG.,
MorschhauserJ.
2000; Activation of the multiple drug resistance gene MDR1 in fluconazole-resistant, clinical Candida albicans strains is caused by mutations in a trans -regulatory factor. J Bacteriol 182:400–404[CrossRef]