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

Bertha Isabel Carvajal-Gamez,Laura Isabel Vázquez-Carrillo,Rossana Arroyo,Laura Itzel Quintas-Granados,Eduardo Carrillo-Tapia,María Elizbeth Alvarez-Sánchez,Lucero De Los Angeles Ramón-Luing,Heidar-Ali Tajmir-Riahi

doi:10.1371/journal.pone.0107293

Bertha Isabel Carvajal-Gamez, Laura Isabel Vázquez-Carrillo + Show 6 more

Open Access

https://doi.org/10.1371/journal.pone.0107293

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Journal: PloS one	Publication Date: Sep 24, 2014
Citations: 48	License type: CC BY 4.0

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.

Highlights

Trichomonosis is the most common non-viral sexually transmitted infection (STI) caused by Trichomonas vaginalis
Since in TvCP65 is involved in trichomonal cytotoxicity and the expression, protein amount, and proteolytic activity of this cysteine proteinase (CP) were reduced in DAB-treated parasites, we suggested that probably exist a relationship between the parasite virulence and polyamines concentration in T. vaginalis [14]
In DAB-treated parasites transferred to exogenous putrescine (DP) (Fig. 1B, lane 3), the TvCP39 activity was,90% restored (Fig. 1C) but not in those parasites transferred into a normal media (DN) (20%) (Fig. 1B, lane 4 and Fig. 1C), suggesting that the activity restoration was due to the exogenous putrescine addition

Summary

Introduction

Trichomonosis is the most common non-viral sexually transmitted infection (STI) caused by Trichomonas vaginalis. This infection mainly affects women, causing vaginitis, cervictis, urethritis, and infertility [1,2] It causes low birth weight infants, preterm delivery [3], and a predisposition to cervical neoplasia [4]. The 39 kDa CP (TvCP39), which was found in vaginal washes from patients with trichomonosis and it is localized in the parasite surface, is involved in cytotoxicity to HeLa, DU145 and vaginal epithelial cells (VECs). This CP is in vivo and in vitro secreted by T. vaginalis, and is active in the pH range found in human vagina and prostate [11]

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