[en] Coccidia are obligate intracellular protozoan parasites responsible for human and veterinary diseases. Eimeria tenella, the aetiologic agent of caecal coccidiosis, is a major pathogen of chickens. In Toxoplasma gondii, some kinases from the rhoptry compartment (ROP) are key virulence factors. ROP kinases hijack and modulate many cellular functions and pathways, allowing T. gondii survival and development. E. tenella's kinome comprises 28 putative members of the ROP kinase family; most of them are predicted, as pseudokinases and their functions have never been characterised. One of the predicted kinase, EtROP1, was identified in the rhoptry proteome of E. tenella sporozoites. Here, we demonstrated that EtROP1 is active, and the N-terminal extension is necessary for its catalytic kinase activity. Ectopic expression of EtROP1 followed by co-immunoprecipitation identified cellular p53 as EtROP1 partner. Further characterisation confirmed the interaction and the phosphorylation of p53 by EtROP1. E. tenella infection or overexpression of EtROP1 resulted both in inhibition of host cell apoptosis and G0/G1 cell cycle arrest. This work functionally described the first ROP kinase from E. tenella and its noncanonical structure. Our study provides the first mechanistic insight into host cell apoptosis inhibition by E. tenella. EtROP1 appears as a new candidate for coccidiosis control.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Diallo, Mamadou Amadou ; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Sausset, Alix; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Gnahoui-David, Audrey; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Ribeiro E Silva, Adeline; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Brionne, Aurélien; BOA, INRA, Université de Tours, Nouzilly, France
Le Vern, Yves; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Bussière, Françoise I; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Tottey, Julie; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Lacroix-Lamandé, Sonia; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Laurent, Fabrice; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
Silvestre, Anne ; Infectiologie et Santé Publique, INRA, Université de Tours, Nouzilly, France
We gratefully acknowledge I. Dimier-Poisson, N. Moiré, and J.F. Dubremetz for providing anti-TgROP2 and anti-SAG1 antibodies. We thank Cassandra Mazaud for the help with in vivo experiment and apoptosis measures in epithelial cell from the caecum from infected chicken. We thank Valérie Labas, Lucie Combes-Soia, and Grégoire Harichaux of the Plate-Forme Chirurgie et Imagerie pour la Recherche et l'Enseignement (Nouzilly, France) for the help with mass spectrometry identification. We thank Luc Reininger for providing us the pGEX-PfTLK (for kinase assay control) and fruitful discussions. We thank Laetitia Fragnet-Trapp (INRA Centre Val de Loire-UMR1282) for helpful discussions on the cell cycle phases. We thank Dr. Mohamed-Ali Hakimi for fruitful discussion on the manuscript.
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