Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts
ABSTRACT Eimeria tenella is an obligate intracellular parasite responsible for avian coccidiosis. Like other apicomplexan parasites, such as Toxoplasma gondii, cell invasion and intracellular development rely on apical organelle content discharge, named micronemes and rhoptries. Some rhoptry (ROP) k...
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American Society for Microbiology
2023-08-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.00137-23 |
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| author | Adeline Ribeiro E. Silva Mamadou Amadou Diallo Alix Sausset Thomas Robert Stéphane Bach Françoise I. Bussière Fabrice Laurent Sonia Lacroix-Lamandé Anne Silvestre |
| author_facet | Adeline Ribeiro E. Silva Mamadou Amadou Diallo Alix Sausset Thomas Robert Stéphane Bach Françoise I. Bussière Fabrice Laurent Sonia Lacroix-Lamandé Anne Silvestre |
| author_sort | Adeline Ribeiro E. Silva |
| collection | DOAJ |
| description | ABSTRACT Eimeria tenella is an obligate intracellular parasite responsible for avian coccidiosis. Like other apicomplexan parasites, such as Toxoplasma gondii, cell invasion and intracellular development rely on apical organelle content discharge, named micronemes and rhoptries. Some rhoptry (ROP) kinases (ROPK) are key virulence factors in T. gondii. To date, among the 28 ropk genes carried by E. tenella, only two to four were confirmed by proteomic analysis or immunostaining to be expressed at the sporozoite stage. We have previously shown that EtROP1 is implicated in the inhibition of host cell apoptosis by interacting with the cellular p53. This work functionally described the second ROP kinase expressed at the sporozoite stage in E. tenella. EtROP2 is an active kinase that phosphorylates cell substrates of approximately 50 kDa. Its overexpression leads to the shortening of the prepatent period and to the early development of first-generation schizonts. Conduction of RNA sequencing analysis and reverse transcriptase quantitative PCR (RT-qPCR) on the host cell allowed us to identify the mitogen-activated protein kinase (MAPK) pathway and the transcription factor cFos to be upregulated by EtROP2. We also showed by immunofluorescence assay that the active kinase EtROP2 is implicated in the p38 MAPK pathway activation. We established here that EtROP2 activates the p38 MAPK pathway through a direct or indirect phosphorylation, leading to the overexpression of the master transcription factor cFos known to be implicated in E. tenella development. IMPORTANCE Rhoptries are specialized secretory organelles found in zoite stages of apicomplexan parasites. In addition to well-conserved rhoptry neck proteins, their protein consists mostly of kinase proteins, highly divergent from eukaryotic kinases. Some of those kinases are described as major virulence factors in Toxoplasma gondii, secreted into the host cell to hijack signaling pathways. Most of those kinases remain to be characterized in Eimeria tenella. Deciphering their cellular function is a prerequisite to supporting their relevance as a druggable target in development of new means of Eimeria tenella control. Secreted divergent kinases that interact with host cell partners to modulate pathways are good candidates, as they coevolve with their host targets to ensure their function within the host and are less prone to mutations that would lead to drug resistance. The absence of any orthologous kinase in host cells makes these parasite kinases a promising drug target candidate. |
| first_indexed | 2024-03-12T14:31:43Z |
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| id | doaj.art-c33d06594168422facd9ec0abe34b316 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-03-12T14:31:43Z |
| publishDate | 2023-08-01 |
| publisher | American Society for Microbiology |
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| series | Microbiology Spectrum |
| spelling | doaj.art-c33d06594168422facd9ec0abe34b3162023-08-17T13:04:14ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972023-08-0111410.1128/spectrum.00137-23Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of SchizontsAdeline Ribeiro E. Silva0Mamadou Amadou Diallo1Alix Sausset2Thomas Robert3Stéphane Bach4Françoise I. Bussière5Fabrice Laurent6Sonia Lacroix-Lamandé7Anne Silvestre8ISP, INRAE, Université de Tours, Nouzilly, FranceISP, INRAE, Université de Tours, Nouzilly, FranceISP, INRAE, Université de Tours, Nouzilly, FranceSorbonne Université, CNRS, UMR 8227, Integrative Biology of Marine Models Laboratory (LBI2M), Station Biologique de Roscoff, Roscoff, FranceSorbonne Université, CNRS, UMR 8227, Integrative Biology of Marine Models Laboratory (LBI2M), Station Biologique de Roscoff, Roscoff, FranceISP, INRAE, Université de Tours, Nouzilly, FranceISP, INRAE, Université de Tours, Nouzilly, FranceISP, INRAE, Université de Tours, Nouzilly, FranceISP, INRAE, Université de Tours, Nouzilly, FranceABSTRACT Eimeria tenella is an obligate intracellular parasite responsible for avian coccidiosis. Like other apicomplexan parasites, such as Toxoplasma gondii, cell invasion and intracellular development rely on apical organelle content discharge, named micronemes and rhoptries. Some rhoptry (ROP) kinases (ROPK) are key virulence factors in T. gondii. To date, among the 28 ropk genes carried by E. tenella, only two to four were confirmed by proteomic analysis or immunostaining to be expressed at the sporozoite stage. We have previously shown that EtROP1 is implicated in the inhibition of host cell apoptosis by interacting with the cellular p53. This work functionally described the second ROP kinase expressed at the sporozoite stage in E. tenella. EtROP2 is an active kinase that phosphorylates cell substrates of approximately 50 kDa. Its overexpression leads to the shortening of the prepatent period and to the early development of first-generation schizonts. Conduction of RNA sequencing analysis and reverse transcriptase quantitative PCR (RT-qPCR) on the host cell allowed us to identify the mitogen-activated protein kinase (MAPK) pathway and the transcription factor cFos to be upregulated by EtROP2. We also showed by immunofluorescence assay that the active kinase EtROP2 is implicated in the p38 MAPK pathway activation. We established here that EtROP2 activates the p38 MAPK pathway through a direct or indirect phosphorylation, leading to the overexpression of the master transcription factor cFos known to be implicated in E. tenella development. IMPORTANCE Rhoptries are specialized secretory organelles found in zoite stages of apicomplexan parasites. In addition to well-conserved rhoptry neck proteins, their protein consists mostly of kinase proteins, highly divergent from eukaryotic kinases. Some of those kinases are described as major virulence factors in Toxoplasma gondii, secreted into the host cell to hijack signaling pathways. Most of those kinases remain to be characterized in Eimeria tenella. Deciphering their cellular function is a prerequisite to supporting their relevance as a druggable target in development of new means of Eimeria tenella control. Secreted divergent kinases that interact with host cell partners to modulate pathways are good candidates, as they coevolve with their host targets to ensure their function within the host and are less prone to mutations that would lead to drug resistance. The absence of any orthologous kinase in host cells makes these parasite kinases a promising drug target candidate.https://journals.asm.org/doi/10.1128/spectrum.00137-23EimeriaApicomplexakinaserhoptryROPKp38 MAPK |
| spellingShingle | Adeline Ribeiro E. Silva Mamadou Amadou Diallo Alix Sausset Thomas Robert Stéphane Bach Françoise I. Bussière Fabrice Laurent Sonia Lacroix-Lamandé Anne Silvestre Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts Microbiology Spectrum Eimeria Apicomplexa kinase rhoptry ROPK p38 MAPK |
| title | Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts |
| title_full | Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts |
| title_fullStr | Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts |
| title_full_unstemmed | Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts |
| title_short | Overexpression of Eimeria tenella Rhoptry Kinase 2 Induces Early Production of Schizonts |
| title_sort | overexpression of eimeria tenella rhoptry kinase 2 induces early production of schizonts |
| topic | Eimeria Apicomplexa kinase rhoptry ROPK p38 MAPK |
| url | https://journals.asm.org/doi/10.1128/spectrum.00137-23 |
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