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Isolation of transcripts overexpressed in the human pathogen Trichophyton rubrum grown in lipid as carbon source
Fernanda C A Maranhão1, Henrique C S Silveira, Antonio Rossi
1Departamento de Genética, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Brazil.
Abstract:
Trichophyton rubrum is the most common etiological agent of human dermatophytosis. Despite the incidence and medical importance of this dermatophyte, little is known about the mechanisms of host invasion and pathogenicity. Host invasion depends on the adaptive cellular responses of the pathogen that allow it to penetrate the skin layers, which are mainly composed of proteins and lipids. In this study, we used suppression subtractive hybridization to identify transcripts overexpressed in T. rubrum cultured in lipid as carbon source. Among the subtractive cDNA clones isolated, 85 clones were positively screened by cDNA array dot blotting and were sequenced. The putative proteins encoded by the isolated transcripts showed similarities to fungal proteins involved in metabolism, signaling, defense, and virulence, such as the MDR/ABC transporter, glucan 1,3-β-glucosidase, chitin synthase B, copper-sulfate-regulated protein, and serine/threonine phosphatase (calcineurin A). These results provide the first molecular insight into the genes differentially expressed during the adaptation of T. rubrum to a lipidic carbon source.
Insights
This study identifies genes in Trichophyton rubrum, a common cause of fungal skin infections, that are active when the fungus uses lipids. This provides insights into how the fungus invades the skin.
Area of Science:
- Medical Mycology
- Molecular Biology
- Biochemistry
Background:
- Trichophyton rubrum is a leading cause of human dermatophytosis.
- Understanding its host invasion and pathogenicity mechanisms is crucial due to its medical importance.
- Skin layers, composed of proteins and lipids, present a significant barrier to fungal penetration.
Purpose of the Study:
- To identify transcripts overexpressed in T. rubrum when utilizing lipids as a carbon source.
- To gain molecular insights into the adaptive cellular responses of T. rubrum during host invasion.
- To uncover genes involved in the pathogenicity of this common dermatophyte.
Main Methods:
- Suppression subtractive hybridization (SSH) was employed to identify differentially expressed genes.
- cDNA array dot blotting was used for positive screening of subtractive clones.
- Sequencing of selected cDNA clones to identify putative protein functions.
Main Results:
- 85 overexpressed transcripts were identified in T. rubrum cultured on lipids.
- Putative proteins showed similarities to fungal enzymes involved in metabolism, signaling, defense, and virulence.
- Identified proteins include MDR/ABC transporter, glucan 1,3-β-glucosidase, chitin synthase B, copper-sulfate-regulated protein, and calcineurin A.
Conclusions:
- This study offers the first molecular perspective on gene expression during T. rubrum's adaptation to a lipidic environment.
- The identified genes are potentially involved in the dermatophyte's ability to penetrate and colonize host skin.
- Further research into these genes could reveal novel targets for antifungal therapies.
