The splicing factor SR2 is an important virulence factor of Toxoplasma gondii
Serine/arginine-rich (SR) proteins are key factors with important roles in constitutive and alternative splicing (AS) of pre-mRNAs. However, the role of SR splicing factors in the pathogenicity of T. gondii remains largely unexplored. Here, we investigated the role of splicing factor SR2, a homolog...
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Frontiers Media S.A.
2023-11-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1302512/full |
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| author | Xiao-Jing Wu Xiao-Jing Wu Jin Gao Jin Gao Xiao-Nan Zheng Hany M. Elsheikha Ting-Ting Li Yong-Jie Kou Yong-Jie Kou Meng Wang Xing-Quan Zhu |
| author_facet | Xiao-Jing Wu Xiao-Jing Wu Jin Gao Jin Gao Xiao-Nan Zheng Hany M. Elsheikha Ting-Ting Li Yong-Jie Kou Yong-Jie Kou Meng Wang Xing-Quan Zhu |
| author_sort | Xiao-Jing Wu |
| collection | DOAJ |
| description | Serine/arginine-rich (SR) proteins are key factors with important roles in constitutive and alternative splicing (AS) of pre-mRNAs. However, the role of SR splicing factors in the pathogenicity of T. gondii remains largely unexplored. Here, we investigated the role of splicing factor SR2, a homolog of Plasmodium falciparum SR1, in the pathogenicity of T. gondii. We functionally characterized the predicted SR2 in T. gondii by gene knockout and studied its subcellular localization by endogenous protein HA tagging using CRISPR-Cas9 gene editing. The results showed that SR2 was localized in the nucleus and expressed in the tachyzoite and bradyzoite stages. In vitro studies including plaque formation, invasion, intracellular replication, egress and bradyzoite differentiation assays showed that deletion of SR2 in type I RH strain and type II Pru strains had no significant effect on the parasite growth and bradyzoite differentiation (p > 0.05). Interestingly, the disruption of SR2 in RH type I (p < 0.0001) and Pru type II (p < 0.05) strains resulted in varying degrees of attenuated virulence. In addition, disruption of SR2 in type II Pru strain significantly reduced brain cyst burden by ~80% (p < 0.0001). Collectively, these results suggest that splicing factor SR2 is important for the pathogenicity of T. gondii, providing a new target for the control and treatment of toxoplasmosis. |
| first_indexed | 2024-03-10T03:11:20Z |
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| id | doaj.art-290a34a47dc34747be56424b19479b7e |
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| issn | 1664-302X |
| language | English |
| last_indexed | 2024-03-10T03:11:20Z |
| publishDate | 2023-11-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Microbiology |
| spelling | doaj.art-290a34a47dc34747be56424b19479b7e2023-11-23T10:23:59ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-11-011410.3389/fmicb.2023.13025121302512The splicing factor SR2 is an important virulence factor of Toxoplasma gondiiXiao-Jing Wu0Xiao-Jing Wu1Jin Gao2Jin Gao3Xiao-Nan Zheng4Hany M. Elsheikha5Ting-Ting Li6Yong-Jie Kou7Yong-Jie Kou8Meng Wang9Xing-Quan Zhu10Laboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province, ChinaState Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, ChinaLaboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province, ChinaState Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, ChinaLaboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province, ChinaFaculty of Medicine and Health Sciences, School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Loughborough, United KingdomState Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, ChinaLaboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province, ChinaState Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, ChinaState Key Laboratory for Animal Disease Control and Prevention, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, ChinaLaboratory of Parasitic Diseases, College of Veterinary Medicine, Shanxi Agricultural University, Taigu, Shanxi Province, ChinaSerine/arginine-rich (SR) proteins are key factors with important roles in constitutive and alternative splicing (AS) of pre-mRNAs. However, the role of SR splicing factors in the pathogenicity of T. gondii remains largely unexplored. Here, we investigated the role of splicing factor SR2, a homolog of Plasmodium falciparum SR1, in the pathogenicity of T. gondii. We functionally characterized the predicted SR2 in T. gondii by gene knockout and studied its subcellular localization by endogenous protein HA tagging using CRISPR-Cas9 gene editing. The results showed that SR2 was localized in the nucleus and expressed in the tachyzoite and bradyzoite stages. In vitro studies including plaque formation, invasion, intracellular replication, egress and bradyzoite differentiation assays showed that deletion of SR2 in type I RH strain and type II Pru strains had no significant effect on the parasite growth and bradyzoite differentiation (p > 0.05). Interestingly, the disruption of SR2 in RH type I (p < 0.0001) and Pru type II (p < 0.05) strains resulted in varying degrees of attenuated virulence. In addition, disruption of SR2 in type II Pru strain significantly reduced brain cyst burden by ~80% (p < 0.0001). Collectively, these results suggest that splicing factor SR2 is important for the pathogenicity of T. gondii, providing a new target for the control and treatment of toxoplasmosis.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1302512/fullToxoplasma gondiiSR proteinssplicing factor SR2alternative splicingpathogenicityCRISPR-Cas9 |
| spellingShingle | Xiao-Jing Wu Xiao-Jing Wu Jin Gao Jin Gao Xiao-Nan Zheng Hany M. Elsheikha Ting-Ting Li Yong-Jie Kou Yong-Jie Kou Meng Wang Xing-Quan Zhu The splicing factor SR2 is an important virulence factor of Toxoplasma gondii Frontiers in Microbiology Toxoplasma gondii SR proteins splicing factor SR2 alternative splicing pathogenicity CRISPR-Cas9 |
| title | The splicing factor SR2 is an important virulence factor of Toxoplasma gondii |
| title_full | The splicing factor SR2 is an important virulence factor of Toxoplasma gondii |
| title_fullStr | The splicing factor SR2 is an important virulence factor of Toxoplasma gondii |
| title_full_unstemmed | The splicing factor SR2 is an important virulence factor of Toxoplasma gondii |
| title_short | The splicing factor SR2 is an important virulence factor of Toxoplasma gondii |
| title_sort | splicing factor sr2 is an important virulence factor of toxoplasma gondii |
| topic | Toxoplasma gondii SR proteins splicing factor SR2 alternative splicing pathogenicity CRISPR-Cas9 |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1302512/full |
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