A neutral trehalase gene from Candida albicans: molecular cloning, characterization and disruption.
A neutral trehalase gene, NTC1, from the human pathogenic yeast Candida albicans was isolated and characterized. An ORF of 2724 bp was identified encoding a predicted protein of 907 amino acids and a molecular mass of 104 kDa. A single transcript of approximately 3.2 kb was detected by Northern blot analysis. Comparison of the deduced amino acid sequence of the C. albicans NTC1 gene product with that of the Saccharomyces cerevisiae NTH1 gene product revealed 57% identity. The NTC1 gene was localized on chromosome 1 or R. A null mutant (delta ntc1/delta ntc1) was constructed by sequential gene disruption. Extracts from mutants homozygous for neutral trehalase deletion had only marginal neutral trehalase activity. Extracts from heterozygous mutants showed intermediate activities between extracts from the wild-type strain and from the homozygous mutants. The null mutant showed no significant differences in pathogenicity as compared to the wild-type strain in a mouse model of systemic candidiasis. This result indicates that the neutral trehalase of C. albicans is not a potential target for antifungal drugs.
Eck
Raimund
R
Hans-KnöUll-Institut für Naturstoff-Forschung e.V., Abteilung Mykologie,Beutenbergstraβe 11, D-07745 Jena,Germany.
Bergmann
Clemens
C
Hans-KnöUll-Institut für Naturstoff-Forschung e.V., Abteilung Mykologie,Beutenbergstraβe 11, D-07745 Jena,Germany.
Ziegelbauer
Karl
K
Bayer AG, PH-R Antiinfectiva Forschung I,D-42096 Wuppertal,Germany.
Schönfeld
Wolfgang
W
Bayer AG, PH-R Antiinfectiva Forschung I,D-42096 Wuppertal,Germany.
Künkel
Waldemer
W
Hans-KnöUll-Institut für Naturstoff-Forschung e.V., Abteilung Mykologie,Beutenbergstraβe 11, D-07745 Jena,Germany.
D000595:Amino Acid Sequence; D000818:Animals; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D002177:Candidiasis / Q000382:microbiology; D002874:Chromosome Mapping; D015825:Chromosomes, Fungal; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D017353:Gene Deletion; D006720:Homozygote; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013712:Tenebrio; D014158:Transcription, Genetic; D014198:Trehalase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*
D011994:Recombinant Proteins; D014198:Trehalase