Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite

• Published in: The journal of microbiology 2017, 55(10), , pp.783-791
• Main Authors: Ospina-Villa, Juan David, Guillén, Nancy, Lopez-Camarillo, Cesar, Soto-Sanchez, Jacqueline, Ramirez-Moreno, Esther, Garcia-Vazquez, Raul, Castañon-Sanchez, Carlos A., Betanzos, Abigail, Marchat, Laurence A.
• Format: Article
• Language: English
• Subjects: Bacteria; Biomedical and Life Sciences; Cell Death; Cell Movement; Cell Proliferation; Cell Survival; Cells; Cleavage; DNA; Double-stranded RNA; Entamoeba histolytica - genetics; Entamoeba histolytica - metabolism; Entamoeba histolytica - pathogenicity; Entamoebiasis - parasitology; Gene expression; Gene Expression Profiling; Gene Expression Regulation; Gene Knockdown Techniques; Gene Silencing; Genes, Protozoan - genetics; Genes, Reporter; Genomics and Molecular Biology; Homology; Human behavior; Humans; Life Sciences; Microbial Genetics; Microbiology; mRNA; mRNA Cleavage and Polyadenylation Factors - genetics; mRNA Cleavage and Polyadenylation Factors - metabolism; Nucleic acids; Nucleotide sequence; Parasite control; Parasites; PCR; Phagocytosis; Polyadenine; Polyadenylation; Polymerase chain reaction; Proliferation; Proteins; Protozoa; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; Ribonucleic acid; RNA; RNA, Messenger - metabolism; RNA, Protozoan - metabolism; Site selection; Trophozoites; Trophozoites - cytology; Trophozoites - metabolism; Virulence Factors - genetics; 생물학
• ISSN: 1225-8873; 1976-3794
• DOI: 10.1007/s12275-017-7259-9

The 25 kDa subunit of the Clevage Factor Im (CFIm25) is an essential factor for messenger RNA polyadenylation in human cells. Therefore, here we investigated whether the homologous protein of Entamoeba histolytica , the protozoan responsible for human amoebiasis, might be considered as a biochemical target for parasite control. Trophozoites were cultured with bacterial double-stranded RNA molecules targeting the EhCFIm25 gene, and inhibition of mRNA and protein expression was confirmed by RT-PCR and Western blot assays, respectively. EhCFIm25 silencing was associated with a significant acceleration of cell proliferation and cell death. Moreover, trophozoites appeared as larger and multinucleated cells. These morphological changes were accompanied by a reduced mobility, and erythrophagocytosis was significantly diminished. Lastly, the knockdown of EhCFIm25 affected the poly(A) site selection in two reporter genes and revealed that EhCFIm25 stimulates the utilization of downstream poly(A) sites in E. histolytica mRNA. Overall, our data confirm that targeting the polyadenylation process represents an interesting strategy for controlling parasites, including E. histolytica . To our best knowledge, the present study is the first to have revealed the relevance of the cleavage factor CFIm25 as a biochemical target in parasites.