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- 저자 : Ce´sar Lo´pez-Camarillo (검색결과 2 건)
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Silencing the cleavage factor CFIm25 as a new strategy to control Entamoeba histolytica parasite
Juan David Ospina-Villa,Nancy Guillén,Ce´sar Lo´pez-Camarillo,Jacqueline Soto-Sanchez,Esther Ramirez-Moreno,Raul Garcia-Vazquez,Carlos A. Castañon-Sanchez,Abigail Betanzos,Laurence A.Marchat 한국미생물학회 2017 The journal of microbiology Vol.55 No.10
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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Transcriptional profile of processing machinery of 30 end of mRNA in Trichomonas vaginalis
Miguel A ´ ngel Del-Moral-Stevenel,Alma Villalobos-Osnaya,Mavil Lo´pez-Casamichana,Laura Itzel Quintas-Granados,Ce´sar Lo´pez-Camarillo,Jose´ Manuel Ferna´ndez Sa´nchez,Selene Zarate-Guerra,Marı´a Eli 한국유전학회 2015 Genes & Genomics Vol.37 No.4
Trichomonas vaginalis is the causative agent of trichomonosis, a sexually transmitted disease (STD) that affects over 180 million people worldwide. This parasite is capable to infect the urogenital tract of women and men, both microenvironments might affect the expression of key genes that may be involved in the parasite pathogenesis. The processing of 30 end of mRNA promotes mRNA stability in many eukaryotes, however in T. vaginalis this molecular machinery is under research. By means of an in silico analysis we identified putative proteins of the 30 end mRNA processing machinery of T. vaginalis, and by RTPCR assays we evaluated the expression of eight of these genes in a female and male T. vaginalis isolates. According to the in silico analysis, the T. vaginalis 30 end mRNA processing machinery, comprises a similar complex and protein factors that those described in Homo sapiens, Arabidopsis thaliana, Saccharomyces cerevisiae and Entamoeba histolytica. The complex contains several subcomplexes, including cleavage and polyadenylation specificity factor (CPSF), cleavage stimulation factor (CstF), cleavage factor I (CFIm) and cleavage factor II (CFIIm). We demonstrated that genes tvpsf2p, tvcfi25, tvcpsf160, tvcpsf73, tvfip1, tvpap1, tvpc4 and tvpabp are expressed in male or female T. vaginalis isolates. Besides we identify two different isoforms of TvPC4. T. vaginalis genome contains most of genes encoding for 30 end mRNA processing, which may be transcriptionally active and could be involved in the capping, splicing, cleavage and polyadenylation of mRNAs in this parasite. Further studies are necessary to elucidate the biological meaning of our findings.
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