Disruption in Candida albicans of the TPS2 gene encoding trehalose-6-phosphate phosphatase affects cell integrity and decreases infectivity.
The gene CaTPS2 encoding trehalose-6-phosphate (T6P) phosphatase from Candida albicans has been cloned and disrupted in this organism. The Catps2/Catps2 mutant did not accumulate trehalose but accumulated high levels of T6P. Disruption of the two copies of the CaTPS2 gene did not abolish growth even at 42 degrees C, but decreased the growth rate. In the stationary phase, the Catps2/Catps2 mutant aggregated, more than 50% of its cells became permeable to propidium iodide and a large amount of protein was found in the culture medium. Aggregation occurred only at pH values higher than 7 and was avoided by osmoprotectants; it was never observed during the exponential phase of growth. The mutant formed colonies with a smooth border on Spider medium. Mice inoculated with 1.5 x 10(6) c.f.u. of wild-type cells died after 8 days, while 80% of those inoculated with the same number of c.f.u. of the Catps2/Catps2 mutant survived for at least 1 month. Reintroduction of the wild-type CaTPS2 gene in the Catps2/Catps2 mutant abolished the phenotypes described. It is hypothesized that the accumulation of T6P interferes with the assembly of a normal cell wall.
Zaragoza
Oscar
O
Instituto de Investigaciones Biomédicas Alberto Sols CSIC-UAM, Unidad de Bioquímica y Genética de Levaduras, 28029 Madrid, Spain.
de Virgilio
Claudio
C
Pontón
José
J
Gancedo
Carlos
C
D000818:Animals; D000935:Antifungal Agents / Q000494:pharmacology; D015139:Blotting, Southern; D002176:Candida albicans / Q000166:cytology* / Q000201:enzymology* / Q000235:genetics / Q000472:pathogenicity; D002177:Candidiasis / Q000382:microbiology; D002473:Cell Wall / Q000648:ultrastructure; D003001:Cloning, Molecular; D017077:Culture Media, Conditioned / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D014774:Virulence
D000935:Antifungal Agents; D017077:Culture Media, Conditioned; C023178:trehalose-phosphatase; D010744:Phosphoric Monoester Hydrolases