León-Sicairos Claudia R, León-Félix Josefina, Arroyo Rossana
Departamento de Patología Experimental, Centro de Investigación y de Estudios Avanzados del IPN (CINVESTAV-IPN), AP 14-740, Mexico City, 07000, Mexico.
Microbiology (Reading). 2004 May;150(Pt 5):1131-1138. doi: 10.1099/mic.0.26927-0.
Trichomonas vaginalis is the causative agent of trichomoniasis, one of the most common sexually transmitted diseases in humans. This protozoan has multiple proteinases that are mainly of the cysteine proteinase (CP) type, some of which are known to be involved in the parasite's virulence. Here, a novel T. vaginalis CP-encoding gene, tvcp12, was identified and characterized. tvcp12 is 948 bp long and encodes a predicted 34.4 kDa protein that has the characteristics of the papain-like CP family. TvCP12 does not appear to have a signal peptide, suggesting that this is a cytoplasmic CP. By Southern blot assays, the tvcp12 gene was found as a single copy in the T. vaginalis genome. Remarkably, Northern blot experiments showed a single transcript band of approximately 1.3 kb in the mRNA obtained from parasites grown in low iron conditions and no transcript was observed in the mRNA from parasites grown in high iron conditions. By RT-PCR assays, a 270 bp band was amplified from the cDNA of parasites grown in low iron medium, which was very faint when cDNA from parasites grown in high iron conditions was used. Transcripts of the 3' region obtained in both iron conditions presented differences in their poly(A) tail length. These data suggest that tvcp12 is another gene that is negatively regulated by iron and that the length of the poly(A) tail may be one of the factors involved in the iron-modulated protein expression.
阴道毛滴虫是滴虫病的病原体,滴虫病是人类最常见的性传播疾病之一。这种原生动物有多种蛋白酶,主要是半胱氨酸蛋白酶(CP)类型,其中一些已知与寄生虫的毒力有关。在此,鉴定并表征了一个新的阴道毛滴虫CP编码基因tvcp12。tvcp12长948 bp,编码一个预测分子量为34.4 kDa的蛋白质,该蛋白质具有木瓜蛋白酶样CP家族的特征。TvCP12似乎没有信号肽,表明这是一种细胞质CP。通过Southern印迹分析,发现tvcp12基因在阴道毛滴虫基因组中为单拷贝。值得注意的是,Northern印迹实验显示,从低铁条件下生长的寄生虫获得的mRNA中有一条约1.3 kb的单转录带,而在高铁条件下生长的寄生虫的mRNA中未观察到转录带。通过RT-PCR分析,从低铁培养基中生长的寄生虫的cDNA中扩增出一条270 bp的条带,而使用高铁条件下生长的寄生虫的cDNA时,该条带非常微弱。在两种铁条件下获得的3'区域转录本在其poly(A)尾长度上存在差异。这些数据表明,tvcp12是另一个受铁负调控的基因,并且poly(A)尾的长度可能是参与铁调节蛋白表达的因素之一。