Isolation of the Candida albicans homologs of Saccharomyces cerevisiae KRE6 and SKN1: expression and physiological function.
Cell wall beta-glucan in a pathogenic fungus, Candida albicans, is highly branched with beta-1,3 and beta-1,6 linkages. We have isolated the C. albicans cDNAs for KRE6 and SKN1, the genes required for beta-1,6-glucan synthesis in Saccharomyces cerevisiae. The results of Northern blot analysis revealed that C. albicans KRE6 was expressed at a higher level than SKN1 in the yeast phase, while SKN1 expression was strongly induced upon induction of hyphal formation. In addition, the C. albicans KRE6 and SKN1 mRNAs but not the actin mRNA were shortened during the yeast-hypha transition. Unlike S. cerevisiae, more than 50% of cell wall glucan was beta-1,6 linked in C. albicans. Neither beta-1,3-glucan nor beta-1,6-glucan was affected by the homozygous C. albicans skn1 delta null mutation. Although we never succeeded in generating the homozygous C. albicans kre6 delta null mutant, the hemizygous kre6 delta mutation decreased the KRE6 mRNA level by about 60% and also caused a more than 80% reduction of beta-1,6-glucan without affecting beta-1,3-glucan. The physiological function of KRE6 was further examined by studying gene regulation in C. albicans. When KRE6 transcription was suppressed by using the HEX1 promoter, C. albicans cells exhibited the partial defect in cell separation and increased susceptibility to Calcofluor White. These results demonstrate that KRE6 plays important roles in beta-1,6-glucan synthesis and budding in C. albicans.
Mio
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Department of Mycology, Nippon Roche Research Center, Kamakura, Japan.
Yamada-Okabe
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Yabe
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Nakajima
T
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Arisawa
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Yamada-Okabe
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H
D000595:Amino Acid Sequence; D001483:Base Sequence; D002176:Candida albicans / Q000235:genetics*; D002473:Cell Wall / Q000378:metabolism; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D005936:Glucans / Q000096:biosynthesis; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D012333:RNA, Messenger / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D013045:Species Specificity; D047071:beta-Glucans
D005656:Fungal Proteins; D005936:Glucans; C081933:KRE6 protein, S cerevisiae; D008565:Membrane Proteins; D012333:RNA, Messenger; C081932:SKN1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D047071:beta-Glucans; C064197:beta-1,6-glucan