After screening of a Candida albicans genome database, the product of an ORF (IPF 3054) that has 62 % homology with Saccharomyces cerevisiae Ssr1p, an internal cell-wall protein, was identified and named CaSsr1p. The deduced amino acid sequence shows that CaSsr1p contains an N-terminal hydrophobic signal peptide, is rich in Ser and Thr amino acids and has a potential glycosylphosphatidylinositol-attachment signal. CaSsr1p is released following degradation of isolated cell walls by zymolyase (mainly a 1,3-β-glucanase) and therefore seems to be covalently linked to the β-glucan of the cell walls. Both disruption and overexpression of the CaSSR1 gene caused an increased sensitivity to calcofluor white, Congo red and zymolyase digestion. These results suggest that CaSsr1p has a structural role associated with the cell-wall β-glucan.
AlaniE.,
CaoL.,
KlecknerN.1987; A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics 116:541–545
BertramG.,
SwobodaR. K.,
GoodayG. W.,
GowN. A. R.,
BrownA. J. P.1996; Structure and regulation of the Candida albicans ADH1 gene encoding an immunogenic alcohol dehydrogenase. Yeast 12:115–127
BoekeJ. D.,
LaCrouteF.,
FinkG. R.1984; A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance. Mol Gen Genet 197:345–346
CarasI. W.,
WeddellG. N.,
DavidzM. A.,
NussenzweigV.,
MartinD. W.Jr1987; Signal for attachment of a phospholipids membrane anchor in decay accelerating factor. Science 238:1280–1283
ElorzaM. V.,
RicoH.,
SentandreuR.1983; Calcofluor white alters the assembly of chitin fibrils in Saccharomyces cerevisiae and Candida albicans cells. J Gen Microbiol 129:1577–1582
ElorzaM. V.,
MurguiA.,
SentandreuR.1985; Dimorphism in Candida albicans : contribution of mannoproteins to the architecture of yeast and mycelial cells. J Gen Microbiol 131:2209–2216
ElorzaM. V.,
MarcillaA.,
SentandreuR.1988; Wall mannoproteins of the yeast and mycelial cells of Candida albicans : nature of the glycosidic bonds and polydispersity of their mannan moieties. J Gen Microbiol 134:2393–2403
GuillumA. M.,
TsayE. Y. H.,
KirschD. R.1984; Isolation of the Candida albicans gene from oritidine-5′-phosphate decarboxylase by complementation of Saccharomyces cerevisiae ura3 and Escherichia coli pyrF mutations. Mol Gen Genet 198:2100–2112
KapteynJ. C.,
Van Den EndeH.,
KlisF. M.1999; The contribution of cell wall proteins to the organization of the yeast cell wall. Biochim Biophys Acta 1426:373–383
KopeckaM.,
GabrielM.1992; The influence of congo red on the cell wall and (1→3)-beta-d-glucan microfibril biogenesis in Saccharomyces cerevisiae . Arch Microbiol 158:115–126
LipkeP. N.,
WojciechowiczD.,
KurjanJ.1989; AGα1 is the structural gene for the Saccharomyces cerevisiae α -agglutinin, a cell surface glycoprotein involved in cell–cell interactions during mating. Mol Cell Biol 9:3155–3165
MonteroM.,
MarcillaA.,
SentandreuR.,
ValentinE.1998; A Candida albicans 37 kDa polypeptide with homology to the laminin receptor is a component of the translational machinery. Microbiology 144:839–847
MontijnR. C.,
van RinsumJ.,
van SchagerF. A.,
KlisF. M.1994; Glucomannoproteins in the cell wall of Saccharomyces cerevisiae contain a novel type of carbohydrate side chain. J Biol Chem 269:19338–19342
MoukadiriI.,
ArmeroJ.,
AbadA.,
SentandreuR.,
ZuecoJ.1997; Identification of a mannoprotein present in the inner layer of the cell wall of Saccharomyces cerevisiae . J Bacteriol 179:2154–2162
MrsaV.,
SeidlT.,
GentzschM.,
TannerW.1997; Specific labelling of cell wall proteins by biotinylation. Identification of four covalently linked O -mannosylated proteins of Saccharomyces cerevisiae . Yeast 13:1145–1154
NakaiK.,
HortonP.1999; psort: a program for detecting sorting signals in proteins and predicting their subcellular localization. Trends Biochem Sci 24:34–36
NielsenH.,
EngelbrechtJ.,
BrunakS.,
von HeijneG.1997; Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10:1–6
NuofferC.,
HorvathA.,
RietzmanH.1993; Analysis of the sequence requirements for glycosylphosphatidylinositol anchoring of Saccharomyces cerevisiae Gas1 protein. J Biol Chem 268:10558–10563
OrleanP.,
AmmerH.,
WatzeleM.,
TannerW.1986; Synthesis of an O -glycosylated cell surface protein induced by alpha factor. Proc Natl Acad Sci U S A 83:6263–6266
PastorF. I. J.,
ValentinE.,
HerreroE.,
SentandreuR.1984; Structure of the Saccharomyces cerevisiae cell wall: mannoproteins released by zymolyase and their contribution to wall architecture. Biochim Biophys Acta 802:292–300
RamA. F. J.,
WoltersA.,
Ten HoopenR.,
KlisF. M.1994; A new approach for isolating cell wall mutants in Saccharomyces cerevisiae by screening for hypersensitivity to calcofluor white. Yeast 10:1019–1130
RamónA. M.,
GilR.,
BurgalM.,
SentandreuR.,
ValentinE.1996; A novel cell wall protein specific to the mycelial form of Yarrowia lipolytica . Yeast 12:1535–1548
RichardM.,
Ibata-OmbettaS.,
DromerF.,
Bordon-PallierF.,
JouaultT.,
GaillardinC.2002; Complete glycosylphosphatidylinositol anchors are required in Candida albicans for full morphogenesis, virulence and resistance to macrophages. Mol Microbiol 44:841–853
SambrookJ.,
FritschE. F.,
ManiatisT.1989Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
SentandreuR.,
ElorzaM. V.,
MormeneoS.,
SanjuanR.,
IranzoM.1993; Possible roles of mannoproteins in the construction of Candida albicans cell wall. In Dimorphic Fungi in Biology and Medicine pp 169–175 Edited by
van den BosscheH.,
OddsF. C.,
KerridgeD.
New York: Plenum;
SentandreuR.,
MormeneoS.,
Ruiz-HerreraJ.1994; Biogenesis of fungal cell wall. In The Mycota I. Growth, Differentiation and Sexuality pp 111–124 Edited by
WesselsJ. G. H.,
MeinhardtF.
Berlin: Springer;
ShermanF.1991; Getting started with yeast. In Guide to Yeast Genetics and Molecular Biology pp 3–21 Edited by
GuthrieC.,
FinkG. R.
San Diego, CA: Academic Press;
TeunissenA. W. R. H.,
HolubE.,
van der HuchtJ.,
van der BergJ. A.,
SteensmaH. Y.1993; Sequence of the open reading frame of the FLO1 gene from Saccharomyces cerevisiae . Yeast 9:423–427
TowbinH.,
StaehelinT.,
GordonJ.1979; Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76:4350–4354
ValentinE.,
HerreroE.,
PastorF. I. J.,
SentandreuR.1984; Solubilization and analysis of mannoprotein molecules from the cell wall of Saccharomyces cerevisiae . J Gen Microbiol 130:1419–1428
ValentinE.,
MormeneoS.,
SentandreuR.2000; The cell surface of Candida albicans during morphogenesis. In Contributions in Microbiology , vol. 5, Dimorphism in Human Pathogenic and Apathogenic Yeasts pp 138–150 Edited by
ErnstJ. F.,
SchmidtA.
Basel: Karger;
van der VaartJ. M.,
CaroL. H.,
ChapmanJ. W.,
KlisF. M.,
VerripsC. T.1995; Identification of three mannoproteins in the cell wall of Saccharomyces cerevisiae . J Bacteriol 177:3104–3110
WojciechowiczD.,
LuC. F.,
KurjanJ.,
LipkeP. N.1993; Cell surface anchoring and ligand-binding domains of the Saccharomyces cerevisiae cell adhesion protein α-agglutinin, a member of the immunoglobulin superfamily. Mol Cell Biol 13:2554–2563