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Microarray and KOG analysis of Acanthamoeba healyi genes up-regulated by mouse-brain passage
Eun-Kyung Moon1, Ying-Hua Xuan2, Hyun-Hee Kong3
1Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, Taegu 700-422, Republic of Korea.
Abstract:
Long-term cultivation in a laboratory could reduce the virulence of Acanthamoeba. To identify virulence factors of Acanthamoeba, the authors compared the transcription profiles of long-term cultivated Acanthamoeba healyi (OLD) and three times mouse-brain passaged A. healyi (MBP) using microarray analysis and eukaryotic orthologous group (KOG) assignments. Microarray analysis revealed that 601 genes were up-regulated by mouse-brain passage. The results of real-time PCR of 8 randomly selected genes up-regulated in the MBP strain confirmed microarray analysis findings. KOG assignments showed relatively higher percentages of the MBP strain up-regulated genes in T article (signal transduction mechanism), O article (posttranslational modification, protein turnover, chaperones), C article (energy production and conversion), and J article (translation, ribosomal structure and biogenesis). In particular, the MBP strain showed higher expressions of cysteine protease and metalloprotease. A comparison of KOG assignments by microarray analysis and previous EST (expressed sequence tags) analysis showed similar populations of up-regulated genes. These results provide important information regarding the identification of virulence factors of pathogenic Acanthamoeba.
Insights
Laboratory cultivation may decrease Acanthamoeba virulence. Mouse-brain passage of Acanthamoeba healyi up-regulated 601 genes, including those for proteases, potentially identifying key virulence factors.
Area of Science:
- Microbiology
- Molecular Biology
- Parasitology
Background:
- Long-term laboratory cultivation can attenuate the virulence of Acanthamoeba species.
- Identifying virulence factors is crucial for understanding and combating Acanthamoeba infections.
Purpose of the Study:
- To compare transcription profiles of laboratory-cultivated and mouse-brain-passaged Acanthamoeba healyi.
- To identify potential virulence factors of Acanthamoeba by analyzing gene expression changes.
Main Methods:
- Microarray analysis was employed to compare gene expression between OLD (long-term cultivated) and MBP (mouse-brain passaged) Acanthamoeba healyi strains.
- Eukaryotic Orthologous Group (KOG) assignments were used to categorize differentially expressed genes.
- Real-time PCR validated the microarray findings for selected up-regulated genes.
Main Results:
- Mouse-brain passage resulted in the up-regulation of 601 genes in A. healyi.
- KOG analysis indicated enrichment of genes involved in signal transduction, posttranslational modification, energy production, and translation in the MBP strain.
- Increased expression of cysteine protease and metalloprotease was observed in the MBP strain.
Conclusions:
- Gene expression profiling effectively identified genes potentially associated with Acanthamoeba virulence.
- Proteases, including cysteine and metalloproteases, are highlighted as potential virulence factors.
- These findings provide valuable insights for future research into pathogenic Acanthamoeba.
