title: Putrescine-dependent re-localization of TvCP39, a cysteine proteinase involved in Trichomonas vaginalis cytotoxicity.
abstract: Polyamines are involved in the regulation of some Trichomonas vaginalis virulence factors such as the transcript, proteolytic activity, and cytotoxicity of TvCP65, a cysteine proteinase (CP) involved in the trichomonal cytotoxicity. In this work, we reported the putrescine effect on TvCP39, other CP that also participate in the trichomonal cytotoxicity. Parasites treated with 1,4-diamino-2-butanone (DAB) (an inhibitor of putrescine biosynthesis), diminished the amount and proteolytic activity of TvCP39 as compared with untreated parasites. Inhibition of putrescine biosynthesis also reduced ∼ 80% the tvcp39 mRNA levels according to RT-PCR and qRT-PCR assays. Additionally, actinomycin D-treatment showed that the tvcp39 mRNA half-life decreased in the absence of putrescine. However, this reduction was restored by exogenous putrescine addition, suggesting that putrescine is necessary for tvcp39 mRNA stability. TvCP39 was localized in the cytoplasm but, in DAB treated parasites transferred into exogenous putrescine culture media, TvCP39 was re-localized to the nucleus and nuclear periphery of trichomonads. Interestingly, the amount and proteolytic activity of TvCP39 was recovered as well as the tvcp39 mRNA levels were restored when putrescine exogenous was added to the DAB-treated parasites. In conclusion, our data show that putrescine regulate the TvCP39 expression, protein amount, proteolytic activity, and cellular localization.
authors.affiliation: Genomic Sciences Postgraduate, Autonomous University of Mexico City (UACM), Mexico City, Mexico.
authors.forename: Bertha Isabel
authors.initials: BI
authors.lastname: Carvajal-Gamez
authors.affiliation: Genomic Sciences Postgraduate, Autonomous University of Mexico City (UACM), Mexico City, Mexico.
authors.forename: Laura Itzel
authors.initials: LI
authors.lastname: Quintas-Granados
authors.affiliation: Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, IPN, Mexico City, Mexico.
authors.forename: Rossana
authors.initials: R
authors.lastname: Arroyo
authors.affiliation: Genomic Sciences Postgraduate, Autonomous University of Mexico City (UACM), Mexico City, Mexico.
authors.forename: Laura Isabel
authors.initials: LI
authors.lastname: Vázquez-Carrillo
authors.affiliation: Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, IPN, Mexico City, Mexico.
authors.forename: Lucero De los Angeles
authors.initials: Lde L
authors.lastname: Ramón-Luing
authors.affiliation: Genomic Sciences Postgraduate, Autonomous University of Mexico City (UACM), Mexico City, Mexico.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Carrillo-Tapia
authors.affiliation: Genomic Sciences Postgraduate, Autonomous University of Mexico City (UACM), Mexico City, Mexico.
authors.forename: María Elizbeth
authors.initials: ME
authors.lastname: Alvarez-Sánchez
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D057056:Cysteine Proteases / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D018613:Microscopy, Confocal; D059748:Proteolysis / Q000187:drug effects; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011700:Putrescine / Q000031:analogs & derivatives / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014246:Trichomonas vaginalis / Q000166:cytology / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; C014458:1,4-diaminobutanone; D057056:Cysteine Proteases; D011700:Putrescine