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Korean J Parasitol. 2017 Apr;55(2):115-120. doi: 10.3347/kjp.2017.55.2.115. Epub 2017 Apr 30.

DNA Methylation of Gene Expression in Acanthamoeba castellanii Encystation.

Author information

1
Department of Medical Zoology, Kyung Hee University School of Medicine, Seoul 02447, Korea.
2
Department of Parasitology and Tropical Medicine, Kyungpook National University School of Medicine, Daegu 41944, Korea.
3
Department of Pharmacology, Kyungpook National University School of Medicine, Daegu 41944, Korea.
4
Department of Parasitology, Dong-A University College of Medicine, Busan 49201, Korea.

Abstract

Encystation mediating cyst specific cysteine proteinase (CSCP) of Acanthamoeba castellanii is expressed remarkably during encystation. However, the molecular mechanism involved in the regulation of CSCP gene expression remains unclear. In this study, we focused on epigenetic regulation of gene expression during encystation of Acanthamoeba. To evaluate methylation as a potential mechanism involved in the regulation of CSCP expression, we first investigated the correlation between promoter methylation status of CSCP gene and its expression. A 2,878 bp of promoter sequence of CSCP gene was amplified by PCR. Three CpG islands (island 1-3) were detected in this sequence using bioinformatics tools. Methylation of CpG island in trophozoites and cysts was measured by bisulfite sequence PCR. CSCP promoter methylation of CpG island 1 (1,633 bp) was found in 8.2% of trophozoites and 7.3% of cysts. Methylation of CpG island 2 (625 bp) was observed in 4.2% of trophozoites and 5.8% of cysts. Methylation of CpG island 3 (367 bp) in trophozoites and cysts was both 3.6%. These results suggest that DNA methylation system is present in CSCP gene expression of Acanthamoeba. In addition, the expression of encystation mediating CSCP is correlated with promoter CpG island 1 hypomethylation.

KEYWORDS:

Acanthamoeba castellanii; encystation; epigenetic regulation

PMID:
28506032
PMCID:
PMC5450953
DOI:
10.3347/kjp.2017.55.2.115
[Indexed for MEDLINE]
Free PMC Article

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