Differential gene expression in the pathogenic dermatophyte Arthroderma benhamiae in vitro versus during infection.
Although dermatophytes are the most common agents of superficial mycoses in humans and animals, the molecular basis of the pathogenicity of these fungi is largely unknown. In vitro digestion of keratin by dermatophytes is associated with the secretion of multiple proteases, which are assumed to be responsible for their particular specialization to colonize and degrade keratinized host structures during infection. To investigate the role of individual secreted proteases in dermatophytosis, a guinea pig infection model was established for the zoophilic dermatophyte Arthroderma benhamiae, which causes highly inflammatory cutaneous infections in humans and rodents. By use of a cDNA microarray covering approximately 20-25 % of the A. benhamiae genome and containing sequences of at least 23 protease genes, we revealed a distinct in vivo protease gene expression profile in the fungal cells, which was surprisingly different from the pattern elicited during in vitro growth on keratin. Instead of the major in vitro -expressed proteases, others were activated specifically during infection. These enzymes are therefore suggested to fulfil important functions that are not exclusively associated with the degradation of keratin. Most notably, the gene encoding the serine protease subtilisin 6, which is a known major allergen in the related dermatophyte Trichophyton rubrum and putatively linked to host inflammation, was found to be the most strongly upregulated gene during infection. In addition, our approach identified other candidate pathogenicity-related factors in A. benhamiae, such as genes encoding key enzymes of the glyoxylate cycle and an opsin-related protein. Our work provides what we believe to be the first broad-scale gene expression profile in human pathogenic dermatophytes during infection, and points to putative virulence-associated mechanisms that make these micro-organisms the most successful aetiological agents of superficial mycoses.
Staib
Peter
P
Leibniz Institute for Natural Product Research and Infection Biology - Hans Knoell Institute, Junior Research Group Fundamental Molecular Biology of Pathogenic Fungi, Beutenbergstr. 11a, D-07745 Jena, Germany.
Zaugg
Christophe
C
Department of Dermatology, Centre Hospitalier Universitaire Vaudois, Av. de Beaumont 29, 1011 Lausanne, Switzerland.
Mignon
Bernard
B
Department of Infectious and Parasitic Diseases, Parasitology, Faculty of Veterinary Medicine, University of Liège, B-43 Sart-Tilman, 4000 Liège, Belgium.
Weber
Johann
J
DNA Array Facility, Center for Integrative Genomics, University of Lausanne, Genopode Building, 1015 Lausanne, Switzerland.
Grumbt
Maria
M
Leibniz Institute for Natural Product Research and Infection Biology - Hans Knoell Institute, Junior Research Group Fundamental Molecular Biology of Pathogenic Fungi, Beutenbergstr. 11a, D-07745 Jena, Germany.
Pradervand
Sylvain
S
DNA Array Facility, Center for Integrative Genomics, University of Lausanne, Genopode Building, 1015 Lausanne, Switzerland.
Harshman
Keith
K
DNA Array Facility, Center for Integrative Genomics, University of Lausanne, Genopode Building, 1015 Lausanne, Switzerland.
Monod
Michel
M
Department of Dermatology, Centre Hospitalier Universitaire Vaudois, Av. de Beaumont 29, 1011 Lausanne, Switzerland.
D000818:Animals; D003883:Arthrodermataceae / Q000201:enzymology / Q000235:genetics*; D003881:Dermatomycoses / Q000382:microbiology*; D005260:Female; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D006168:Guinea Pigs; D007633:Keratins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism*; D012331:RNA, Fungal / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D012331:RNA, Fungal; D007633:Keratins; D010447:Peptide Hydrolases