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Characterization of the Neospora caninum NcROP40 and NcROP2Fam-1 rhoptry proteins during the tachyzoite lytic cycle

Published online by Cambridge University Press:  02 November 2015

IVÁN PASTOR-FERNÁNDEZ Open the ORCID record for IVÁN PASTOR-FERNÁNDEZ [Opens in a new window] ,
JAVIER REGIDOR-CERRILLO ,
ELENA JIMÉNEZ-RUIZ ,
GEMA ÁLVAREZ-GARCÍA ,
VIRGINIA MARUGÁN-HERNÁNDEZ ,
ANDREW HEMPHILL  and
LUIS M. ORTEGA-MORA
IVÁN PASTOR-FERNÁNDEZ
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain
JAVIER REGIDOR-CERRILLO
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain
ELENA JIMÉNEZ-RUIZ
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain
GEMA ÁLVAREZ-GARCÍA
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain
VIRGINIA MARUGÁN-HERNÁNDEZ
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain
ANDREW HEMPHILL
Affiliation:
Institute of Parasitology, Vetsuisse Faculty, University of Berne, Länggass-Strasse 122, CH-3012 Berne, Switzerland
LUIS M. ORTEGA-MORA*
Affiliation:
SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain
*
* Corresponding author: SALUVET, Animal Health Department, Faculty of Veterinary Sciences, Complutense University of Madrid, Ciudad Universitaria s/n, 28040-Madrid, Spain. E-mail: luis.ortega@vet.ucm.es
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Summary

Virulence factors from the ROP2-family have been extensively studied in Toxoplasma gondii, but in the closely related Neospora caninum only NcROP2Fam-1 has been partially characterized to date. NcROP40 is a member of this family and was found to be more abundantly expressed in virulent isolates. Both NcROP2Fam-1 and NcROP40 were evaluated as vaccine candidates and exerted a synergistic effect in terms of protection against vertical transmission in mouse models, which suggests that they may be relevant for parasite pathogenicity. NcROP40 is localized in the rhoptry bulbs of tachyzoites and bradyzoites, but in contrast to NcROP2Fam-1, the protein does not associate with the parasitophorous vacuole membrane due to the lack of arginine-rich amphipathic helix in its sequence. Similarly to NcROP2Fam-1, NcROP40 mRNA levels are highly increased during tachyzoite egress and invasion. However, NcROP40 up-regulation does not appear to be linked to the mechanisms triggering egress. In contrast to NcROP2Fam-1, phosphorylation of NcROP40 was not observed during egress. Besides, NcROP40 secretion into the host cell was not successfully detected by immunofluorescence techniques. These findings indicate that NcROP40 and NcROP2Fam-1 carry out different functions, and highlight the need to elucidate the role of NcROP40 within the lytic cycle and to explain its relative abundance in tachyzoites.

Keywords

Neospora caninumNcROP40NcROP2Fam-1characterizationin silico analysislytic cycle of tachyzoitesimmunolocalizationsecretion assaysmRNA expression profileprotein phosphorylation

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Type
Research Article
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Parasitology , Volume 143 , Issue 1 , January 2016 , pp. 97 - 113
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Copyright © Cambridge University Press 2015 

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