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Disruption in Candida albicans of the TPS2 gene encoding trehalose-6-phosphate phosphatase affects cell integrity and decreases infectivity

Instituto de Investigaciones Biomédicas ‘Alberto Sols’ CSIC-UAM, Unidad de Bioquímica y Genética de Levaduras, 28029 Madrid, Spain¹
Botanisches Institut, Universität Basel, Hebelstr 1, CH-4056 Basel, Switzerland²
Departamento de Inmunología, Microbiología y Parasitología, Facultad de Medicina y Odontología, Universidad del País Vasco, 48080 Bilbao, Spain³

Author for correspondence: Carlos Gancedo. Tel: +34 91 5854620. Fax: +34 91 5854587. e-mail: cgancedo{at}iib.uam.es

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 °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 1·5x10⁶ 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.

Keywords: cell wall, aggregation, stress, antifungals

Abbreviations: T6P, trehalose 6-phosphate

The EMBL accession number for the sequence reported in this paper is AJ242990.

^a Present address: Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461, USA.

^b Present address: Département de Biochimie Médicale, Centre Médical Universitaire, 1211 Geneva 4, Switzerland.

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