title: The knockdown of each component of the cysteine proteinase-adhesin complex of Entamoeba histolytica (EhCPADH) affects the expression of the other complex element as well as the in vitro and in vivo virulence.
abstract: Entamoeba histolytica is the protozoan parasite causative of human amoebiasis, disease responsible for 40 000-100 000 deaths annually. The cysteine proteinase-adhesin complex of this parasite (EhCPADH) is a heterodimeric protein formed by a cysteine protease (EhCP112) and an adhesin (EhADH) that plays an important role in the cytopathic mechanism of this parasite. The coding genes for EhCP112 and EhADH are adjacent in the E. histolytica genome, suggesting that their expression may be co-regulated, but this hypothesis has not yet been confirmed. Here, we performed the knockdown of EhCP112 and EhADH using gene-specific short-hairpin RNAs (shRNA), and the effect of these knockdowns on the expression of both complex components as well as on the in vitro and in vivo virulence was analysed. Results showed that the knockdown of one of the EhCPADH components produced a simultaneous downregulation of the other protein. Accordingly, a concomitant reduction in the overall expression of the complex was observed. The downregulation of each component also produced a significant decrease in the in vitro and in vivo virulence of trophozoites. These results demonstrated that the expression of EhCP112 and EhADH is co-regulated and confirmed that the EhCPADH complex plays an important role in E. histolytica virulence.
authors.affiliation: Departamento de Infectómica y Patogénesis Molecular,CINVESTAV-IPN,México,D.F. México.
authors.forename: Ramón
authors.initials: R
authors.lastname: Ocádiz-Ruiz
authors.affiliation: Departamento de Infectómica y Patogénesis Molecular,CINVESTAV-IPN,México,D.F. México.
authors.forename: Wendy
authors.initials: W
authors.lastname: Fonseca
authors.affiliation: Department of Biochemistry and Molecular Genetics,University of Virginia Health System,Charlottesville,Virginia 22908,USA.
authors.forename: Alicia S
authors.initials: AS
authors.lastname: Linford
authors.affiliation: Department of Pathobiological Sciences,School of Veterinary Medicine,University of Wisconsin,Madison,Wisconsin 53706,USA.
authors.forename: Timothy P
authors.initials: TP
authors.lastname: Yoshino
authors.affiliation: Departamento de Infectómica y Patogénesis Molecular,CINVESTAV-IPN,México,D.F. México.
authors.forename: Esther
authors.initials: E
authors.lastname: Orozco
authors.affiliation: Departamento de Infectómica y Patogénesis Molecular,CINVESTAV-IPN,México,D.F. México.
authors.forename: Mario A
authors.initials: MA
authors.lastname: Rodríguez
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D006224:Cricetinae; D057056:Cysteine Proteases / Q000235:genetics* / Q000378:metabolism; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity; D004749:Entamoebiasis / Q000469:parasitology*; D005260:Female; D005786:Gene Expression Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D046912:Multiprotein Complexes; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D053058:Trophozoites / Q000378:metabolism; D014774:Virulence
chemical_list: D000913:Antibodies, Protozoan; D046912:Multiprotein Complexes; D015800:Protozoan Proteins; D034741:RNA, Small Interfering; D057056:Cysteine Proteases
keywords: EhCPADH complex; Entamoeba histolytica; RNA interference; downregulation; expression co-regulation; shRNA; virulence