beta-(1,3)-glucan synthase complex from Alternaria infectoria, a rare dematiaceous human pathogen

Anjos, J.; Fernandes, C.; Silva, B. M. A.; Quintas, Célia; Abrunheiro, A.; Gow, N. A. R.; Gonçalves, Teresa

http://hdl.handle.net/10400.1/7303

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The fungal cell wall polymer beta-(1,3)-D-glucan is synthesized by the enzyme beta-(1,3)D-glucan synthase that is a complex composed of at least two proteins, Rho1p and Fks1p. Here, we report the nucleotide sequence of a single FKS gene and of the regulatory unit, RHO1 from the dematiaceous pathogenic fungus Alternaria infectoria. The predicted AiFks and AiRho share, respectively, 93% and 100% identity with that of Drechslera tritici-repentis. We also report that the sensitivity to caspofungin of eight different A. infectoria clinical strains is similar, with a MIC > 32 mu g/ml and a MEC of 1 mu g/ml, except for one strain which had a MEC of 1.4 mu g/ml. This same strain exhibited one substitution at the hot spot 2, S1405A, compatible with less susceptible phenotypes, with the other seven strains having no mutations in either hot spot 1 or 2. The relative quantification of the expression of AiFKS and of AiRHO demonstrated a decrease in response to an exposure to caspofungin at 0.5 mu g/ml.