GAO Xue Juan; FENG Jun Xia; ZHU Sen; LIU Xiao Hui; TARDIEUX Isabelle; LIU Lang Xia

doi:10.3967/bes2014.130

doi: 10.3967/bes2014.130

基金项目: the Natural Science Foundation of Guangdong Province(Grant 9151065004000005)%National Program on Key Basic Research Project (973 Program)(Grant 2011CB910700)%High-Level Talents Project of the Universities of Guangdong([2011]431)%National Natural Science Foundation of China(Grant 31000628,81071790)%Natural Science Foundation of Guangdong Province(Grant S2013030013315)%Fundamental Research Funds for the Central Universities(Grant 21611430,21610101,21609317)

Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, Jinan University, Guangzhou 510632, Guangdong, China
Institut Cochin, Université Descartes, Team FRM‘Barriers and Pathogens’, 22 Rue Méchain, Paris 75014, France

Funds: the Natural Science Foundation of Guangdong Province(Grant 9151065004000005)%National Program on Key Basic Research Project (973 Program)(Grant 2011CB910700)%High-Level Talents Project of the Universities of Guangdong([2011]431)%National Natural Science Foundation of China(Grant 31000628,81071790)%Natural Science Foundation of Guangdong Province(Grant S2013030013315)%Fundamental Research Funds for the Central Universities(Grant 21611430,21610101,21609317)

摘要: Objective The protozoan Toxoplasma gondii expresses large amounts of a 37 kDa Type 2C serine-threonine phosphatase, the so-called TgPP2C which has been suggested to contribute to parasite growth regulation. Ectopic expression in mammalian cells also indicated that the enzyme could regulate growth and survival. In this study, we aimed to investigate the interaction of TgPP2C with human SSRP1 (structure-specific recognition protein 1) and the effects of TgPP2C on cell viability.
Methods The yeast two hybrid system, His-tag pull-down and co-immunoprecipitation assays were used to confirm the interaction of TgPP2C with SSRP1 and determine the binding domain on SSRP1. The evaluation of cell apoptosis was performed using cleaved caspase-3 antibody and Annexin-V/PI kit combined with flow cytometry.
Results We identified human SSRP1 as an interacting partner of TgPP2C. The C-terminal region of SSRP1 including the amino acids 471 to 538 was specifically mapped as the region responsible for interaction with TgPP2C. The overexpression of TgPP2C down-regulated cell apoptosis and negatively regulated apoptosis induced by DRB, casein kinase II (CKII) inhibitor, through enhanced interaction with SSRP1.
Conclusion TgPP2C may be a parasitic factor capable of promoting cell survival through interaction with the host protein SSRP1, thereby creating a favorable environment for parasite growth.
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Abstract: Objective The protozoan Toxoplasma gondii expresses large amounts of a 37 kDa Type 2C serine-threonine phosphatase, the so-called TgPP2C which has been suggested to contribute to parasite growth regulation. Ectopic expression in mammalian cells also indicated that the enzyme could regulate growth and survival. In this study, we aimed to investigate the interaction of TgPP2C with human SSRP1 (structure-specific recognition protein 1) and the effects of TgPP2C on cell viability.
Methods The yeast two hybrid system, His-tag pull-down and co-immunoprecipitation assays were used to confirm the interaction of TgPP2C with SSRP1 and determine the binding domain on SSRP1. The evaluation of cell apoptosis was performed using cleaved caspase-3 antibody and Annexin-V/PI kit combined with flow cytometry.
Results We identified human SSRP1 as an interacting partner of TgPP2C. The C-terminal region of SSRP1 including the amino acids 471 to 538 was specifically mapped as the region responsible for interaction with TgPP2C. The overexpression of TgPP2C down-regulated cell apoptosis and negatively regulated apoptosis induced by DRB, casein kinase II (CKII) inhibitor, through enhanced interaction with SSRP1.
Conclusion TgPP2C may be a parasitic factor capable of promoting cell survival through interaction with the host protein SSRP1, thereby creating a favorable environment for parasite growth.
- Apoptosis /
- SSRP1 /
- TgPP2C /
- Toxoplasma gondii

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doi: 10.3967/bes2014.130

Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

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Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

doi: 10.3967/bes2014.130

English Abstract

Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, Jinan University, Guangzhou 510632, Guangdong, China

Institut Cochin, Université Descartes, Team FRM‘Barriers and Pathogens’, 22 Rue Méchain, Paris 75014, France

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doi: 10.3967/bes2014.130

Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

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Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

doi: 10.3967/bes2014.130

English Abstract

Protein Phosphatase 2C of Toxoplasma Gondii Interacts with Human SSRP1 and Negatively Regulates Cell Apoptosis

Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, Jinan University, Guangzhou 510632, Guangdong, China Institut Cochin, Université Descartes, Team FRM‘Barriers and Pathogens’, 22 Rue Méchain, Paris 75014, France

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目录

Key Laboratory of Functional Protein Research of Guangdong Higher Education Institutes, Institute of Life and Health Engineering, Jinan University, Guangzhou 510632, Guangdong, China

Institut Cochin, Université Descartes, Team FRM‘Barriers and Pathogens’, 22 Rue Méchain, Paris 75014, France