Comprehensive characterization of purine and pyrimidine transport activities in Trichomonas vaginalis and functional cloning of a trichomonad nucleoside transporter,Molecular Microbiology

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Comprehensive characterization of purine and pyrimidine transport activities in Trichomonas vaginalis and functional cloning of a trichomonad nucleoside transporter
Molecular Microbiology ( IF 2.6 ) Pub Date : 2021-11-04 , DOI: 10.1111/mmi.14840
Manal J Natto ₁ , Yukiko Miyamoto ₂ , Jane C Munday ₁ , Tahani A AlSiari ₁ , Mohammed I Al-Salabi ₁ , Neils B Quashie ₁ , Anthonius A Eze ₁ , Lars Eckmann ₂ , Harry P De Koning ₁

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Trichomoniasis is a common and widespread sexually-transmitted infection, caused by the protozoan parasite Trichomonas vaginalis. T. vaginalis lacks the biosynthetic pathways for purines and pyrimidines, making nucleoside metabolism a drug target. Here we report the first comprehensive investigation into purine and pyrimidine uptake by T. vaginalis. Multiple carriers were identified and characterized with regard to substrate selectivity and affinity. For nucleobases, a high-affinity adenine transporter, a possible guanine transporter and a low affinity uracil transporter were found. Nucleoside transporters included two high affinity adenosine/guanosine/uridine/cytidine transporters distinguished by different affinities to inosine, a lower affinity adenosine transporter, and a thymidine transporter. Nine Equilibrative Nucleoside Transporter (ENT) genes were identified in the T. vaginalis genome. All were expressed equally in metronidazole-resistant and -sensitive strains. Only TvagENT2 was significantly upregulated in the presence of extracellular purines; expression was not affected by co-culture with human cervical epithelial cells. All TvagENTs were cloned and separately expressed in Trypanosoma brucei. We identified the main broad specificity nucleoside carrier, with high affinity for uridine and cytidine as well as purine nucleosides including inosine, as TvagENT3. The in-depth characterization of purine and pyrimidine transporters provides a critical foundation for the development of new anti-trichomonal nucleoside analogues.

中文翻译：

阴道毛滴虫嘌呤和嘧啶转运活性的综合表征及毛滴虫核苷转运蛋白的功能克隆

滴虫病是一种常见且广泛的性传播感染，由原生动物寄生虫阴道毛滴虫引起。阴道毛滴虫缺乏嘌呤和嘧啶的生物合成途径，使得核苷代谢成为药物靶标。在此，我们报告了首次对阴道毛滴虫摄取嘌呤和嘧啶的全面研究。鉴定了多种载体并对其底物选择性和亲和力进行了表征。对于核碱基，发现了高亲和力腺嘌呤转运蛋白、可能的鸟嘌呤转运蛋白和低亲和力尿嘧啶转运蛋白。核苷转运蛋白包括两种高亲和力腺苷/鸟苷/尿苷/胞苷转运蛋白，其区别在于对肌苷的不同亲和力、低亲和力腺苷转运蛋白和胸苷转运蛋白。在阴道毛滴虫基因组中鉴定出九个平衡核苷转运蛋白 (ENT) 基因。所有这些在甲硝唑耐药菌株和敏感菌株中表达相同。在细胞外嘌呤存在的情况下，只有 TvagENT2 显着上调；与人宫颈上皮细胞共培养不影响表达。所有 TvagENT 均在布氏锥虫中克隆并单独表达。我们确定了主要的广泛特异性核苷载体，即 TvagENT3，对尿苷和胞苷以及包括肌苷在内的嘌呤核苷具有高亲和力。嘌呤和嘧啶转运蛋白的深入表征为新型抗滴虫核苷类似物的开发提供了关键基础。

更新日期：2021-12-20

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