title: cDNA representational difference analysis used in the identification of genes expressed by Trichophyton rubrum during contact with keratin.
abstract: Dermatophytes are adapted to infect skin, hair and nails by their ability to utilize keratin as a nutrient source. Trichophyton rubrum is an anthropophilic fungus, causing up to 90% of chronic cases of dermatophytosis. The understanding of the complex interactions between the fungus and its host should include the identification of genes expressed during infection. To identify the genes involved in the infection process, representational difference analysis (RDA) was applied to two cDNA populations from T. rubrum, one transcribed from the RNA of fungus cultured in the presence of keratin and the other from RNA generated during fungal growth in minimal medium. The analysis identified differentially expressed transcripts. Genes related to signal transduction, membrane protein, oxidative stress response, and some putative virulence factors were up-regulated during the contact of the fungus with keratin. The expression patterns of these genes were also verified by real-time PCR, in conidia of T. rubrum infecting primarily cultured human keratinocytes in vitro, revealing their potential role in the infective process. A better understanding of this interaction will contribute significantly to our knowledge of the process of dermatophyte infection.
authors: Laboratório de Micologia Clínica, Departamento de Análises Clínicas, Faculdade de Ciências Farmacêuticas, UNESP, CEP 14801-902, 1621 Araraquara, SP, Brazil.
authors: Lilian Cristiane
authors: LC
authors: Baeza
authors: Alexandre Melo
authors: AM
authors: Bailão
authors: Clayton Luiz
authors: CL
authors: Borges
authors: Maristela
authors: M
authors: Pereira
authors: Célia Maria de Almeida
authors: CM
authors: Soares
authors: Maria José Soares
authors: MJ
authors: Mendes Giannini
mesh_terms: D015152:Blotting, Northern; D002478:Cells, Cultured; D003470:Culture Media; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism*; D020224:Expressed Sequence Tags; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D006801:Humans; D015603:Keratinocytes / Q000382:microbiology; D007633:Keratins / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D014249:Trichophyton / Q000145:classification / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity
chemical_list: D003470:Culture Media; D018076:DNA, Complementary; D005656:Fungal Proteins; D012331:RNA, Fungal; D012333:RNA, Messenger; D007633:Keratins