Schmidt Juliana C, Manhães Lauro, Fragoso Stenio P, Pavoni Daniela P, Krieger Marco A
Laboratory of Functional Genomics, Instituto Carlos Chagas, Oswaldo Cruz Foundation (FIOCRUZ), Curitiba, Paraná, Brazil; Health Science Department, Universidade Comunitária da Região de Chapecó (UNOCHAPECÓ), Chapecó, Santa Catarina, Brazil.
Laboratory of Functional Genomics, Instituto Carlos Chagas, Oswaldo Cruz Foundation (FIOCRUZ), Curitiba, Paraná, Brazil.
Parasitol Int. 2018 Apr;67(2):131-139. doi: 10.1016/j.parint.2017.10.009. Epub 2017 Nov 20.
The protozoan Trypanosoma cruzi is a parasite exposed to several environmental stressors inside its invertebrate and vertebrate hosts. Although stress conditions are involved in its differentiation processes, little information is available about the stress response proteins engaged in these activities. This work reports the first known association of the stress-inducible protein 1 (STI1) with the cellular differentiation process in a unicellular eukaryote. Albeit STI1 expression is constitutive in epimastigotes and metacyclic trypomastigotes, higher protein levels were observed in late growth phase epimastigotes subjected to nutritional stress. Analysis by indirect immunofluorescence revealed that T. cruzi STI1 (TcSTI1) is located throughout the cell cytoplasm, with some cytoplasmic granules appearing in greater numbers in late growing epimastigotes and late growing epimastigotes subjected to nutritional stress. We observed that part of the fluorescence signal from both TcSTI1 and TcHSP70 colocalized around the nucleus. Gene silencing of sti1 in Trypanosoma brucei did not affect cell growth. Similarly, the growth of T. cruzi mutant parasites with a single allele sti1 gene knockout was not affected. However, the differentiation of epimastigotes in metacyclic trypomastigotes (metacyclogenesis) was compromised. Lower production rates and numbers of metacyclic trypomastigotes were obtained from the mutant parasites compared with the wild-type parasites. These data indicate that reduced levels of TcSTI1 decrease the rate of in vitro metacyclogenesis, suggesting that this protein may participate in the differentiation process of T. cruzi.
原生动物克氏锥虫是一种寄生虫,在其无脊椎动物和脊椎动物宿主体内会面临多种环境应激源。尽管应激条件参与其分化过程,但关于参与这些活动的应激反应蛋白的信息却很少。这项工作报道了应激诱导蛋白1(STI1)与单细胞真核生物细胞分化过程的首次已知关联。尽管STI1在无鞭毛体和循环后期锥鞭毛体中呈组成性表达,但在受到营养应激的生长后期无鞭毛体中观察到更高的蛋白水平。间接免疫荧光分析显示,克氏锥虫STI1(TcSTI1)位于整个细胞质中,在生长后期无鞭毛体和受到营养应激的生长后期无鞭毛体中,一些细胞质颗粒数量更多。我们观察到来自TcSTI1和TcHSP70的部分荧光信号在细胞核周围共定位。布氏锥虫中sti1的基因沉默不影响细胞生长。同样,单等位基因sti1基因敲除的克氏锥虫突变寄生虫的生长也不受影响。然而,无鞭毛体向循环后期锥鞭毛体的分化(循环后期发育)受到损害。与野生型寄生虫相比,突变寄生虫产生的循环后期锥鞭毛体的生产率和数量更低。这些数据表明,TcSTI1水平降低会降低体外循环后期发育的速率,表明该蛋白可能参与克氏锥虫的分化过程。