Toxoplasma gondii ROP18: potential to manipulate host cell mitochondrial apoptosis
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Toxoplasma gondii is an obligate intracellular parasite that may manipulate host cell mitochondrial apoptosis pathways. In our experiment, 293T cells were transfected with the p3×FLAG-CMV-Myc-ROP18 vector and expressed the ROP18-Myc fusion protein. Cell apoptosis was induced by 0.5 μg/mL actinomycin D (ActD) and was detected by Annexin V-FITC/PI assay. The cell mitochondrial membrane potential was determined by JC-1. Cytochrome c (Cyto-c) from mitochondria and the cytoplasm was measured by Western blot. The Bcl-2 and Bax coding gene expression levels were detected by real-time PCR. We found, in vitro, that T. gondii ROP18 significantly suppressed 293T cell apoptosis induced by ActD and maintained mitochondrial membrane potential and integrity, thereby preventing the release of Cyto-c from mitochondria into the cytoplasm. The ratio of Bcl-2/Bax in ROP18-overexpressing cells was significantly higher than that of the negative control. Therefore, we speculate that ROP18 could suppress host cell apoptosis via the mitochondrial apoptosis pathway in vitro.
KeywordsToxoplasma gondii ROP18 Apoptosis Mitochondrial apoptosis pathway
This study received financial support from the National Natural Science Foundation of China (grant number 81301453), the Laboratory of Parasite and Vector Biology of China, MOPH (grant number WSBKTKT201302), the China Postdoctoral Science Special Foundation(grant number 2015T80518), the China Postdoctoral Science Foundation (grant number 2014M561598), Jiangsu Postdoctoral Science Foundation (grant number 1402171C), and the Senior Talent Studying Initial Funding of Jiangsu University(grant number 13JDG023, 13JDG127).
- Hippe D, Weber A, Zhou L, Chang DC, Hacker G, Luder CG (2009) Toxoplasma gondii infection confers resistance against BimS-induced apoptosis by preventing the activation and mitochondrial targeting of pro-apoptotic Bax. J Cell Sci 122(Pt19):3511–3521. doi: 10.1242/jcs.050963 CrossRefPubMedGoogle Scholar
- Marsden VS, O’Connor L, O’Reilly LA, Silke J, Metcalf D, Ekert PG, Huang DC, Cecconi F, Kuida K, Tomaselli KJ, Roy S, Nicholson DW, Vaux DL, Bouillet P, Adams JM, Strasser A (2002) Apoptosis initiated by Bcl-2-regulated caspase activation independently of the cytochrome c/Apaf-1/caspase-9 apoptosome. Nature 419(6907):634–637CrossRefPubMedGoogle Scholar