- Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite
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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
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J. Microbiol. 2017;55(10):783-791. Published online September 28, 2017
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DOI: https://doi.org/10.1007/s12275-017-7259-9
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 Abstract
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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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Citations
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