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expressed throughout sporulation of the apicomplexan parasite Eimeria tenella
Anaïs Rieux, Simon Gras, Fabien Lecaille, Alisson Niepceron, Marilyn Katrib, Nicholas C. Smith, Gilles Lalmanach, Fabien Brossier
To cite this version:
Anaïs Rieux, Simon Gras, Fabien Lecaille, Alisson Niepceron, Marilyn Katrib, et al.. Eimeripain,
a cathepsin B-like cysteine protease, expressed throughout sporulation of the apicomplexan para-
site Eimeria tenella. PLoS ONE, Public Library of Science, 2012, 7 (3), pp.e31914. �10.1371/jour-
nal.pone.0031914�. �hal-02648245�
Eimeripain, a Cathepsin B-Like Cysteine Protease,
Expressed throughout Sporulation of the Apicomplexan Parasite Eimeria tenella
Anaı¨s Rieux
1,2¤, Simon Gras
1,2, Fabien Lecaille
3, Alisson Niepceron
1,2, Marilyn Katrib
4, Nicholas C. Smith
5, Gilles Lalmanach
3, Fabien Brossier
1,2*
1
INRA, UMR1282, Equipe Pathogene`se des Coccidioses, Infectiologie et Sante´ Publique, Nouzilly, France,
2Universite´ Franc¸ois Rabelais de Tours, UMR1282, Infectiologie et Sante´ Publique, Tours, France,
3INSERM U618, Prote´ases et Vectorisation Pulmonaires, Universite´ Franc¸ois Rabelais, Tours, France,
4Institute for the Biotechnology of Infectious Diseases, University of Technology, Sydney, Australia,
5Queensland Tropical Health Alliance, Faculty of Medicine, Health and Molecular Sciences, James Cook University, Cairns, Australia
Abstract
The invasion and replication of Eimeria tenella in the chicken intestine is responsible for avian coccidiosis, a disease that has major economic impacts on poultry industries worldwide. E. tenella is transmitted to naı¨ve animals via shed unsporulated oocysts that need contact with air and humidity to form the infectious sporulated oocysts, which contain the first invasive form of the parasite, the sporozoite. Cysteine proteases (CPs) are major virulence factors expressed by protozoa. In this study, we show that E. tenella expresses five transcriptionally regulated genes encoding one cathepsin L, one cathepsin B and three cathepsin Cs. Biot-LC-LVG-CHN
2, a cystatin derived probe, tagged eight polypeptides in unsporulated oocysts but only one in sporulated oocysts. CP-dependant activities were found against the fluorescent substrates, Z-FR-AMC and Z-LR- AMC, throughout the sporulation process. These activities corresponded to a cathepsin B-like enzyme since they were inhibited by CA-074, a specific cathepsin B inhibitor. A 3D model of the catalytic domain of the cathepsin B-like protease, based on its sequence homology with human cathepsin B, further confirmed its classification as a papain-like protease with similar characteristics to toxopain-1 from the related apicomplexan parasite, Toxoplasma gondii; we have, therefore, named the E. tenella cathepsin B, eimeripain. Following stable transfection of E. tenella sporozoites with a plasmid allowing the expression of eimeripain fused to the fluorescent protein mCherry, we demonstrated that eimeripain is detected throughout sporulation and has a punctate distribution in the bodies of extra- and intracellular parasites. Furthermore, CA- 074 Me, the membrane-permeable derivative of CA-074, impairs invasion of epithelial MDBK cells by E. tenella sporozoites.
This study represents the first characterization of CPs expressed by a parasite from the Eimeria genus. Moreover, it emphasizes the role of CPs in transmission and dissemination of exogenous stages of apicomplexan parasites.
Citation:
Rieux A, Gras S, Lecaille F, Niepceron A, Katrib M, et al. (2012) Eimeripain, a Cathepsin B-Like Cysteine Protease, Expressed throughout Sporulation of the Apicomplexan Parasite
Eimeria tenella. PLoS ONE 7(3): e31914. doi:10.1371/journal.pone.0031914Editor:
John Parkinson, Hospital for Sick Children, Canada
Received
October 21, 2011;
AcceptedJanuary 17, 2012;
PublishedMarch 22, 2012
Copyright:ß
2012 Rieux et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding:
Funding came from the French National Institute for Agricultural Research. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing Interests: