| Autor: |
Ospina-Villa JD; Instituto Politécnico Nacional - ENMH, Ciudad de México, Mexico., Guillén N; Institut Pasteur, Unité d'Analyses d'Images Biologiques, Paris, France., Lopez-Camarillo C; Universidad Autónoma de la Ciudad de México - Posgrado en Ciencias Genómicas, Ciudad de México, Mexico., Soto-Sanchez J; Instituto Politécnico Nacional - ENMH, Ciudad de México, Mexico., Ramirez-Moreno E; Instituto Politécnico Nacional - ENMH, Ciudad de México, Mexico., Garcia-Vazquez R; Instituto Politécnico Nacional - ENMH, Ciudad de México, Mexico., Castañon-Sanchez CA; Hospital Regional de Alta Especialidad, Oaxaca, Mexico., Betanzos A; Cátedras, CONACYT, Departamento de Infectómica y Patogénesis Molecular, CINVESTAV-IPN, Ciudad de México, Mexico., Marchat LA; Instituto Politécnico Nacional - ENMH, Ciudad de México, Mexico. lmarchat@gmail.com. |
| Abstrakt: |
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. |
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