title: Secreted metalloprotease gene family of Microsporum canis.
abstract: Keratinolytic proteases secreted by dermatophytes are likely to be virulence-related factors. Microsporum canis, the main agent of dermatophytosis in dogs and cats, causes a zoonosis that is frequently reported. Using Aspergillus fumigatus metalloprotease genomic sequence (MEP) as a probe, three genes (MEP1, MEP2, and MEP3) were isolated from an M. canis genomic library. They presented a quite-high percentage of identity with both A. fumigatus MEP and Aspergillus oryzae neutral protease I genes. At the amino acid level, they all contained an HEXXH consensus sequence, confirming that these M. canis genes (MEP genes) encode a zinc-containing metalloprotease gene family. Furthermore, MEP3 was found to be the gene encoding a previously isolated M. canis 43.5-kDa keratinolytic metalloprotease, and was successfully expressed as an active recombinant enzyme in Pichia pastoris. Reverse transcriptase nested PCR performed on total RNA extracted from the hair of M. canis-infected guinea pigs showed that at least MEP2 and MEP3 are produced during the infection process. This is the first report describing the isolation of a gene family encoding potential virulence-related factors in dermatophytes.
authors.affiliation: Department of Parasitology and Parasitic Diseases, Faculty of Veterinary Medicine, University of Liège, Liège, Belgium.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Brouta
authors.forename: Frédéric
authors.initials: F
authors.lastname: Descamps
authors.forename: Michel
authors.initials: M
authors.lastname: Monod
authors.forename: Sandy
authors.initials: S
authors.lastname: Vermout
authors.forename: Bertrand
authors.initials: B
authors.lastname: Losson
authors.forename: Bernard
authors.initials: B
authors.lastname: Mignon
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001232:Aspergillus fumigatus / Q000201:enzymology / Q000235:genetics; D001236:Aspergillus oryzae / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D002415:Cats; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D003881:Dermatomycoses / Q000209:etiology; D004285:Dogs; D005656:Fungal Proteins; D015870:Gene Expression; D005800:Genes, Fungal; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D008864:Microsporum / Q000201:enzymology* / Q000235:genetics* / Q000472:pathogenicity; D008969:Molecular Sequence Data; D005810:Multigene Family; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D014774:Virulence / Q000235:genetics / Q000502:physiology; D015047:Zoonoses / Q000209:etiology
chemical_list: D004271:DNA, Fungal; D005656:Fungal Proteins; D011994:Recombinant Proteins; C117855:MEP1 protein, Haemonchus contortus; D008666:Metalloendopeptidases