{"title":"弓形虫ROP17通过Bcl-2-Beclin 1途径促进自噬。","authors":"Min Guo, Jia Sun, Wen-Tao Wang, Hong-Yan Liu, Yue-Hua Liu, Ke-Ru Qin, Jin-Rui Hu, Xin-Yang Li, Hong-Li Liu, Wei Wang, Zhao-Yang Chen, Chun-Fang Wang, Hai-Long Wang","doi":"10.14411/fp.2021.016","DOIUrl":null,"url":null,"abstract":"<p><p>The apicomplexan Toxoplasma gondii (Nicolle et Manceaux, 1908) secretes a group of serine/threonine kinases from rhoptries, which play vital roles in boosting intracellular infection. Toxoplasma gondii rhoptry organelle protein 17 (ROP17) is one of these important kinase proteins. Nevertheless, its function remains unclear. Here, we showed that ROP17 induced autophagy in vitro and in vivo. The autophagy of small intestine tissues of T. gondii tachyzoite (RH strain)-infected mice was detected by the immunohistochemistry staining of LC3B, Beclin 1 and P62. ROP17 overexpression augmented starvation-induced autophagy in HEK 293T cells as measured by MDC staining, transmission electron microscopy (TEM), fluorescence microscopy and Western blot analysis. Moreover, the interaction of ROP17 and Bcl-2 was confirmed using co-immunoprecipitation analysis, and the data demonstrated that ROP17 had an autophagic role dependent on the Beclin 1-Bcl-2 pathway, which was also revealed in an in vivo model through immunohistochemical staining. Pearson coefficient analysis showed that there existed strong positive correlations between the expression of ROP17 and LC3B, Beclin 1 and phosphorylation of Bcl-2, while strong negative correlations between the expression of ROP17 and p62 and Bcl-2. Collectively, our findings indicate that ROP17 plays a pivotal role in maintaining T. gondii proliferation in host cells via the promotion of autophagy-dependent survival.</p>","PeriodicalId":55154,"journal":{"name":"Folia Parasitologica","volume":null,"pages":null},"PeriodicalIF":1.5000,"publicationDate":"2021-06-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"2","resultStr":"{\"title\":\"Toxoplasma gondii ROP17 promotes autophagy via the Bcl-2-Beclin 1 pathway.\",\"authors\":\"Min Guo, Jia Sun, Wen-Tao Wang, Hong-Yan Liu, Yue-Hua Liu, Ke-Ru Qin, Jin-Rui Hu, Xin-Yang Li, Hong-Li Liu, Wei Wang, Zhao-Yang Chen, Chun-Fang Wang, Hai-Long Wang\",\"doi\":\"10.14411/fp.2021.016\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The apicomplexan Toxoplasma gondii (Nicolle et Manceaux, 1908) secretes a group of serine/threonine kinases from rhoptries, which play vital roles in boosting intracellular infection. Toxoplasma gondii rhoptry organelle protein 17 (ROP17) is one of these important kinase proteins. Nevertheless, its function remains unclear. Here, we showed that ROP17 induced autophagy in vitro and in vivo. The autophagy of small intestine tissues of T. gondii tachyzoite (RH strain)-infected mice was detected by the immunohistochemistry staining of LC3B, Beclin 1 and P62. ROP17 overexpression augmented starvation-induced autophagy in HEK 293T cells as measured by MDC staining, transmission electron microscopy (TEM), fluorescence microscopy and Western blot analysis. Moreover, the interaction of ROP17 and Bcl-2 was confirmed using co-immunoprecipitation analysis, and the data demonstrated that ROP17 had an autophagic role dependent on the Beclin 1-Bcl-2 pathway, which was also revealed in an in vivo model through immunohistochemical staining. Pearson coefficient analysis showed that there existed strong positive correlations between the expression of ROP17 and LC3B, Beclin 1 and phosphorylation of Bcl-2, while strong negative correlations between the expression of ROP17 and p62 and Bcl-2. Collectively, our findings indicate that ROP17 plays a pivotal role in maintaining T. gondii proliferation in host cells via the promotion of autophagy-dependent survival.</p>\",\"PeriodicalId\":55154,\"journal\":{\"name\":\"Folia Parasitologica\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2021-06-18\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"2\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Folia Parasitologica\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.14411/fp.2021.016\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"PARASITOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Folia Parasitologica","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.14411/fp.2021.016","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"PARASITOLOGY","Score":null,"Total":0}
引用次数: 2
摘要
顶端复合体弓形虫(Nicolle et Manceaux, 1908)从弓形虫体内分泌一组丝氨酸/苏氨酸激酶,这些激酶在促进细胞内感染中起着至关重要的作用。弓形虫弓形体细胞器蛋白17 (ROP17)是这些重要的激酶蛋白之一。然而,它的功能仍不清楚。本研究表明,在体外和体内,ROP17均能诱导自噬。采用LC3B、Beclin 1和P62免疫组化染色检测弓形虫速殖子(RH株)感染小鼠小肠组织的自噬。通过MDC染色、透射电镜(TEM)、荧光显微镜和Western blot分析发现,ROP17过表达增强了HEK 293T细胞饥饿诱导的自噬。此外,通过共免疫沉淀分析证实了ROP17与Bcl-2的相互作用,数据表明ROP17具有依赖于Beclin 1-Bcl-2通路的自噬作用,通过免疫组织化学染色在体内模型中也揭示了这一点。Pearson系数分析显示,ROP17与LC3B、Beclin 1的表达与Bcl-2磷酸化呈强正相关,而ROP17与p62的表达与Bcl-2呈强负相关。总之,我们的研究结果表明,ROP17通过促进自噬依赖性存活,在维持弓形虫在宿主细胞中的增殖中起着关键作用。
Toxoplasma gondii ROP17 promotes autophagy via the Bcl-2-Beclin 1 pathway.
The apicomplexan Toxoplasma gondii (Nicolle et Manceaux, 1908) secretes a group of serine/threonine kinases from rhoptries, which play vital roles in boosting intracellular infection. Toxoplasma gondii rhoptry organelle protein 17 (ROP17) is one of these important kinase proteins. Nevertheless, its function remains unclear. Here, we showed that ROP17 induced autophagy in vitro and in vivo. The autophagy of small intestine tissues of T. gondii tachyzoite (RH strain)-infected mice was detected by the immunohistochemistry staining of LC3B, Beclin 1 and P62. ROP17 overexpression augmented starvation-induced autophagy in HEK 293T cells as measured by MDC staining, transmission electron microscopy (TEM), fluorescence microscopy and Western blot analysis. Moreover, the interaction of ROP17 and Bcl-2 was confirmed using co-immunoprecipitation analysis, and the data demonstrated that ROP17 had an autophagic role dependent on the Beclin 1-Bcl-2 pathway, which was also revealed in an in vivo model through immunohistochemical staining. Pearson coefficient analysis showed that there existed strong positive correlations between the expression of ROP17 and LC3B, Beclin 1 and phosphorylation of Bcl-2, while strong negative correlations between the expression of ROP17 and p62 and Bcl-2. Collectively, our findings indicate that ROP17 plays a pivotal role in maintaining T. gondii proliferation in host cells via the promotion of autophagy-dependent survival.
期刊介绍:
FOLIA PARASITOLOGICA, issued in online versions, is an international journal that covers the whole field of general, systematic, ecological and experimental parasitology. It publishes original research papers, research notes and review articles. Contributions from all branches of animal parasitology, such as morphology, taxonomy, biology, biochemistry, physiology, immunology, molecular biology and evolution of parasites, and host-parasite relationships, are eligible. Novelty and importance in the international (not local or regional) context are required. New geographical records of parasites, records of new hosts, regional parasite and/or host surveys (if they constitute the principal substance of manuscript), local/regional prevalence surveys of diseases, local/regional studies on epidemiology of well known diseases and of parasite impact on human/animal health, case reports, routine clinical studies and testing of established diagnostic or treatment procedures, will not be considered. One species description will also not be considered unless they include more general information, such as new diagnostic characters, host-parasite associations, phylogenetic implications, etc. Manuscripts found suitable on submission will be reviewed by at least two reviewers.