DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species
Abstract Background Toxoplasma gondii is an obligate parasite of all warm-blooded animals around the globe. Once infecting a cell, it manipulates the host’s DNA damage response that is yet to be elucidated. The objectives of the present study were three-fold: (i) to assess DNA damages in T. gondii-i...
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doaj-bcb4f777d3da4ab5af52b1545aaa56f02020-11-25T03:55:48ZengBMCParasites & Vectors1756-33052020-09-011311910.1186/s13071-020-04324-7DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen speciesHaohan Zhuang0Chaoqun Yao1Xianfeng Zhao2Xueqiu Chen3Yimin Yang4Siyang Huang5Lingtao Pan6Aifang Du7Yi Yang8Institute of Preventive Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang UniversityDepartments of Biomedical Sciences and One Health Center for Zoonoses and Tropical Veterinary Medicine, Ross University School of Veterinary MedicineAnimals & Plant Inspection and Quarantine Technology Center of Shenzhen CustomsInstitute of Preventive Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang UniversityInstitute of Preventive Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang UniversityInstitute of Comparative Medicine, College of Veterinary Medicine, Yangzhou University, and Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonosis, and Jiangsu Key Laboratory of ZoonosisInstitute of Preventive Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang UniversityInstitute of Preventive Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang UniversityInstitute of Preventive Veterinary Medicine, Zhejiang Provincial Key Laboratory of Preventive Veterinary Medicine, College of Animal Sciences, Zhejiang UniversityAbstract Background Toxoplasma gondii is an obligate parasite of all warm-blooded animals around the globe. Once infecting a cell, it manipulates the host’s DNA damage response that is yet to be elucidated. The objectives of the present study were three-fold: (i) to assess DNA damages in T. gondii-infected cells in vitro; (ii) to ascertain causes of DNA damage in T. gondii-infected cells; and (iii) to investigate activation of DNA damage responses during T. gondii infection. Methods HeLa, Vero and HEK293 cells were infected with T. gondii at a multiplicity of infection (MOI) of 10:1. Infected cells were analyzed for a biomarker of DNA double-strand breaks (DSBs) γH2AX at 10 h, 20 h or 30 h post-infection using both western blot and immunofluorescence assay. Reactive oxygen species (ROS) levels were measured using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), and ROS-induced DNA damage was inhibited by a ROS inhibitor N-acetylcysteine (NAC). Lastly, DNA damage responses were evaluated by detecting the active form of ataxia telangiectasia mutated/checkpoint kinase 2 (ATM/CHK2) by western blot. Results γH2AX levels in the infected HeLa cells were significantly increased over time during T. gondii infection compared to uninfected cells. NAC treatment greatly reduced ROS and concomitantly diminished γH2AX in host cells. The phosphorylated ATM/CHK2 were elevated in T. gondii-infected cells. Conclusions Toxoplasma gondii infection triggered DNA DSBs with ROS as a major player in host cells in vitro. It also activated DNA damage response pathway ATM/CHK2. Toxoplasma gondii manages to keep a balance between survival and apoptosis of its host cells for the benefit of its own survival.http://link.springer.com/article/10.1186/s13071-020-04324-7Toxoplasma gondiiDNA damageReactive oxygen speciesDNA damage response |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Haohan Zhuang Chaoqun Yao Xianfeng Zhao Xueqiu Chen Yimin Yang Siyang Huang Lingtao Pan Aifang Du Yi Yang |
spellingShingle |
Haohan Zhuang Chaoqun Yao Xianfeng Zhao Xueqiu Chen Yimin Yang Siyang Huang Lingtao Pan Aifang Du Yi Yang DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species Parasites & Vectors Toxoplasma gondii DNA damage Reactive oxygen species DNA damage response |
author_facet |
Haohan Zhuang Chaoqun Yao Xianfeng Zhao Xueqiu Chen Yimin Yang Siyang Huang Lingtao Pan Aifang Du Yi Yang |
author_sort |
Haohan Zhuang |
title |
DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species |
title_short |
DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species |
title_full |
DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species |
title_fullStr |
DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species |
title_full_unstemmed |
DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species |
title_sort |
dna double-strand breaks in the toxoplasma gondii-infected cells by the action of reactive oxygen species |
publisher |
BMC |
series |
Parasites & Vectors |
issn |
1756-3305 |
publishDate |
2020-09-01 |
description |
Abstract Background Toxoplasma gondii is an obligate parasite of all warm-blooded animals around the globe. Once infecting a cell, it manipulates the host’s DNA damage response that is yet to be elucidated. The objectives of the present study were three-fold: (i) to assess DNA damages in T. gondii-infected cells in vitro; (ii) to ascertain causes of DNA damage in T. gondii-infected cells; and (iii) to investigate activation of DNA damage responses during T. gondii infection. Methods HeLa, Vero and HEK293 cells were infected with T. gondii at a multiplicity of infection (MOI) of 10:1. Infected cells were analyzed for a biomarker of DNA double-strand breaks (DSBs) γH2AX at 10 h, 20 h or 30 h post-infection using both western blot and immunofluorescence assay. Reactive oxygen species (ROS) levels were measured using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), and ROS-induced DNA damage was inhibited by a ROS inhibitor N-acetylcysteine (NAC). Lastly, DNA damage responses were evaluated by detecting the active form of ataxia telangiectasia mutated/checkpoint kinase 2 (ATM/CHK2) by western blot. Results γH2AX levels in the infected HeLa cells were significantly increased over time during T. gondii infection compared to uninfected cells. NAC treatment greatly reduced ROS and concomitantly diminished γH2AX in host cells. The phosphorylated ATM/CHK2 were elevated in T. gondii-infected cells. Conclusions Toxoplasma gondii infection triggered DNA DSBs with ROS as a major player in host cells in vitro. It also activated DNA damage response pathway ATM/CHK2. Toxoplasma gondii manages to keep a balance between survival and apoptosis of its host cells for the benefit of its own survival. |
topic |
Toxoplasma gondii DNA damage Reactive oxygen species DNA damage response |
url |
http://link.springer.com/article/10.1186/s13071-020-04324-7 |
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