Putrescine-dependent re-localization of TvCP39, a cysteine proteinase involved in Trichomonas vaginalis cytotoxicity

• Published in: PloS one Vol. 9; no. 9; p. e107293
• Main Authors: Carvajal-Gamez, Bertha Isabel, Quintas-Granados, Laura Itzel, Arroyo, Rossana, Vázquez-Carrillo, Laura Isabel, Ramón-Luing, Lucero De los Angeles, Carrillo-Tapia, Eduardo, Alvarez-Sánchez, María Elizbeth
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 24-09-2014; Public Library of Science (PLoS)
• Subjects: Actinomycin; Active Transport, Cell Nucleus - drug effects; Biology and life sciences; Biosynthesis; Blotting, Western; Cell culture; Cell Division - drug effects; Cell Nucleus - drug effects; Cell Nucleus - metabolism; Culture media; Cysteine; Cysteine Proteases - genetics; Cysteine Proteases - metabolism; Cysteine proteinase; Cytoplasm; Cytotoxicity; Entamoeba histolytica; Gene expression; Gene Expression - drug effects; Infections; Localization; Media (culture); Metabolism; Metabolites; Microscopy, Confocal; mRNA stability; Nuclei; Parasites; Parasitic diseases; Parasitology; Pathogenesis; Polyamines; Polymerase chain reaction; Proteinase; Proteins; Proteolysis; Proteolysis - drug effects; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; Putrescine; Putrescine - analogs & derivatives; Putrescine - antagonists & inhibitors; Putrescine - metabolism; Putrescine - pharmacology; Reverse Transcriptase Polymerase Chain Reaction; Sexually transmitted diseases; STD; Stem cells; Toxicity; Transcription; Trichomonas vaginalis; Trichomonas vaginalis - cytology; Trichomonas vaginalis - genetics; Trichomonas vaginalis - metabolism; Virulence; Virulence factors; Mexico
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0107293

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.