Proteomic profile of Wangiella dermatitidis /

Abstract

Wangiella dermatitidis is a pathogenic fungus capable of expressing three distinct modes of cellular development. These forms include yeast, multicellular, and mould phases. In particular, the latter two morphologies are induced from the yeast phase by specific environmental conditions. Temperature-sensitive mutants, designated Mc strains, have also been derived that grow as yeasts at 25°C, but develop in the multicellular phenotype when incubated at 37°C. The latter strongly resembles that tissue form of certain pathogenic fungi phylogenetically related to W dermatitidis. These observations form the basis ofthe following hypothesis: the conversion from the yeastlike budding phase to the multicellular growth pattern is associated with the expression of specific proteins. Therefore, the present study seeks to establish a proteome map ofW dermatitidis for subsequent studies targeting virulence-associated proteins. Yeast-phase cultures ofW dermatitidis were incubated for 24 hours at 25°C and 37°C prior to homogenization. Protein components of the homogenate were resolved by two-dimensional gel electrophoresis, then analyzed by fluorescence imaging using Sypro Ruby Stain. A proteomic profile was created for the wild-type strain 8656 at both temperatures as well as for the mutant strain Mc3. Distinct protein spots separated in the profiles were excised and are being further identified by mass spectrometry. The collective results ofthe present study demonstrate the potential of standard proteomic techniques for dissecting the molecular mechanisms of virulence in W dermatitidis.