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

Juan David Ospina-Villa; Nancy Guillén; Cesar Lopez-Camarillo; Jacqueline Soto-Sanchez; Esther Ramirez-Moreno; Raul Garcia-Vazquez; Carlos A. Castañon-Sanchez; Abigail Betanzos; Laurence A. Marchat

doi:10.1007/s12275-017-7259-9

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Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite: Juan David Ospina-Villa ¹, Nancy Guillén ², Cesar Lopez-Camarillo ³, Jacqueline Soto-Sanchez ¹, Esther Ramirez-Moreno ¹, Raul Garcia-Vazquez ¹, Carlos A. Castañon-Sanchez ⁴, Abigail Betanzos ⁵, Laurence A. Marchat ¹; Journal of Microbiology 2017;55(10):783-791.
DOI: https://doi.org/10.1007/s12275-017-7259-9
Published online: September 28, 2017

¹Instituto Politécnico Nacional – ENMH, Ciudad de México, Mexico, ²Institut Pasteur, Unité d’Analyses d’Images Biologiques, Paris, France, ³Universidad Autónoma de la Ciudad de México – Posgrado en Ciencias Genómicas, Ciudad de México, Mexico, ⁴Hospital Regional de Alta Especialidad, Oaxaca, Mexico, ⁵Cátedras, CONACYT, Departamento de Infectómica y Patogénesis Molecular, CINVESTAV-IPN, Ciudad de México, Mexico¹Instituto Politécnico Nacional – ENMH, Ciudad de México, Mexico, ²Institut Pasteur, Unité d’Analyses d’Images Biologiques, Paris, France, ³Universidad Autónoma de la Ciudad de México – Posgrado en Ciencias Genómicas, Ciudad de México, Mexico, ⁴Hospital Regional de Alta Especialidad, Oaxaca, Mexico, ⁵Cátedras, CONACYT, Departamento de Infectómica y Patogénesis Molecular, CINVESTAV-IPN, Ciudad de México, Mexico

Corresponding author: Laurence A. Marchat , Tel: +52-55 5729-6300 (ext. 55543),

Received: 27 June 2017 • Revised: 16 August 2017 • Accepted: 19 August 2017

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Abstract

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.

Keywords: amoebiasis;gene knockdown;polyadenylation;protozoan parasite;virulence

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

J. Microbiol. 2017;55(10):783-791. Published online September 28, 2017

DOI: https://doi.org/10.1007/s12275-017-7259-9

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