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Published on: September 13, 2022
Identifying differentially expressed genes in the dimorphic fungus Penicillium marneffei by suppression subtractive
Hongfang Liu1, Liyan Xi, Junmin Zhang
1Department of Dermatology, The Second Affiliated Hospital, Sun Yat-Sen University, Guangzhou, Guangdong, China.
Abstract:
Penicillium marneffei is the only known Penicillium species that exhibits temperature-dependent dimorphic growth. It usually infects immunocompromised individuals and may cause fatal penicilliosis marneffei. Its pathogenicity appears to be intimately related to the dimorphic transition from a mycelial phase in the environment to a yeast phase in the human host. To further understand the molecular mechanisms in the switching process of Penicillium marneffei, differential gene expression analysis was carried out between the mycelial phase and the yeast phase by suppression subtractive hybridization. Five hundred genes from forward and reverse-subtracted cDNA libraries were screened by cDNA array dot blotting and 43 genes were identified to be differentially expressed. These genes were found to be homologs of genes involved in various cellular processes including cell wall synthesis, signal transduction, cell cycle, substance transport, general metabolism and stress response, etc. Six genes were further analyzed using relative quantification real-time reverse transcriptase PCR. It is believed that these genes determine some of the major characteristics of dimorphism in Penicillium marneffei.
Insights
Penicillium marneffei switches between mold and yeast forms, causing disease in immunocompromised people. Researchers identified 43 differentially expressed genes critical for this temperature-dependent dimorphic transition.
Area of Science:
- Mycology
- Medical Mycology
- Molecular Biology
Background:
- Penicillium marneffei is unique for its temperature-dependent dimorphism.
- This fungus causes fatal penicilliosis in immunocompromised individuals.
- Dimorphic transition is key to its pathogenicity, switching from mycelial to yeast form.
Purpose of the Study:
- To investigate the molecular mechanisms underlying the dimorphic transition in Penicillium marneffei.
- To identify genes involved in the switch between mycelial and yeast phases.
Main Methods:
- Differential gene expression analysis using suppression subtractive hybridization.
- Screening of 500 genes via cDNA array dot blotting.
- Validation of 6 key genes using real-time reverse transcriptase PCR.
Main Results:
- Identified 43 differentially expressed genes between mycelial and yeast phases.
- These genes are homologs involved in cell wall synthesis, signal transduction, cell cycle, transport, metabolism, and stress response.
- Six specific genes were further analyzed for their role in dimorphism.
Conclusions:
- The identified differentially expressed genes are crucial for the dimorphic characteristics of Penicillium marneffei.
- Understanding these genes provides insights into the pathogenicity of P. marneffei infections.
- This study lays the groundwork for further research into controlling dimorphism and related diseases.
