Project description:The enteric protozoan parasite Entamoeba histolytica have high phagocytic ability. Phagocytosis is also important for the pathogenicity of this parasite; molecular mechanisms of phagocytosis and phagosome maturation is focused. Atg8 is well studied autophagy marker protein. We previously shown that E. histolytica Atg8 translocate to nascent trogosomes and expression silencing of the Atg8 caused retardation of phagosome acidification. To investigate how Atg8 regulates phagosome maturation, here we conducted proteome analysis of phagosomes isolated from an E. histolytica strain in which atg8 gene expression are silenced (atg8 gene silenced, atg8-gs) and its mock control strain (transfected with psAP2-Gunma).
Project description:Entamoeba histolytica trophozoites epigenetically silenced for ameobapore A (G3 parasites) were additionally silenced for the Rhomboid gene Rom1 (EHI_197460) using the mechanism outlined in Mirelman et al. Parasite. 2008 Sep;15(3):266-74 (PMID 18814693). Rom1 silencing was confirmed by RT-PCR. Gene expression in these parasites was compared to that of the parent G3 strain.
Project description:Entamoeba histolytica trophozoites epigenetically silenced for ameobapore A (G3 parasites) were additionally silenced for the Rhomboid gene Rom1 (EHI_197460) using the mechanism outlined in Mirelman et al. Parasite. 2008 Sep;15(3):266-74 (PMID 18814693). Rom1 silencing was confirmed by RT-PCR. Gene expression in these parasites was compared to that of the parent G3 strain. RNA was isolated from axenically grown G3 and Rom1 silenced parasites, and hybridized to a custom Affymetrix array for E. histolytica. Two biological replicates were performed for each condition.
Project description:Previously, we have shown that in Entamoeba histolytica, methionine γ-lyase (EhMGL) downregulation results in trifluoromethionine resistance. The transcriptional response, however, of this parasite to the drug is not known. In this study, we used microarray analysis to determine if additional genes are involved. The expression profile of 9,230 genes in wild type and trifluoromethionine resistant strains was compared. Episomal overexpression of EhBspA1 was performed to verify its role in trifluoromethionine resistance. The transcriptomes of trifluoromethionine resistant strain cultured with or without trifluoromethionine, EhMGL-gene silenced strain, metronidazole resistant strain, and wild type strain under cysteine-deprived conditions were compared to determine the specificity of the changes observed in trifluoromethionine resistant strain.
Project description:To know the role of lysosomal hydrolase receptors in virulecne of enteric protozoan parasite Entamoeba histolytica, eleven cysteine protease binding protein family (CPBF) proteins were specifically silenced and examined the effect on virulence-related phenotypes. Among them, CPBF2 gene silence caused defect in Matrigel invasion. To clarify transcriptomic difference in CPBF2 gene silenceing strain, RNA-seq analysis was conducted.
