TY - JOUR
T1 - Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite
AU - Ospina-Villa, Juan David
AU - Guillén, Nancy
AU - Lopez-Camarillo, Cesar
AU - Soto-Sanchez, Jacqueline
AU - Ramirez-Moreno, Esther
AU - Garcia-Vazquez, Raul
AU - Castañon-Sanchez, Carlos A.
AU - Betanzos, Abigail
AU - Marchat, Laurence A.
N1 - Publisher Copyright:
© 2017, The Microbiological Society of Korea and Springer-Verlag GmbH Germany.
PY - 2017/10/1
Y1 - 2017/10/1
N2 - 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.
AB - 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.
KW - amoebiasis
KW - gene knockdown
KW - polyadenylation
KW - protozoan parasite
KW - virulence
UR - http://www.scopus.com/inward/record.url?scp=85029959406&partnerID=8YFLogxK
U2 - 10.1007/s12275-017-7259-9
DO - 10.1007/s12275-017-7259-9
M3 - Artículo
C2 - 28956353
SN - 1225-8873
VL - 55
SP - 783
EP - 791
JO - Journal of Microbiology
JF - Journal of Microbiology
IS - 10
ER -