Cieśla Joanna, Jagielska Elzbieta, Skopiński Tomasz, Abrowska Magdalena D, Maley Frank, Rode Wojciech
Department of Cellular Biochemistry, Nencki Institute of Experimental Biology, 3 Pasteur St., 02-093 Warsaw, Poland.
Biochem J. 2005 Sep 15;390(Pt 3):681-8. doi: 10.1042/BJ20050548.
Thymidylate synthase (TS) of Trichinella spiralis, a parasitic nematode causing trichinellosis, was found to bind its own mRNA and repress translation of the latter, similar to its human counter-part [Chu, Koeller, Casey, Drake, Chabner, Elwood, Zinn and Allegra (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 8977-8981]. However, in striking contrast with human TS, the parasite enzyme's interaction with mRNA was not affected by any of the substrate (deoxyuridylate or N(5,10)-methylenetetrahydrofolate) nor by the inhibitor (fluorodeoxyuridylate; used alone or in the presence of N(5,10)-methylenetetrahydrofolate) similar to that shown for the bifunctional enzyme from Plasmodium falciparum [Zhang and Rathod (2002) Science 296, 545-547]. Moreover, repression of the translation of the parasite enzyme was enhanced by the same ligands that were shown by others (Chu et al., 1991) to prevent human TS from impairing its translation. On comparing the capacity of TS to bind to its cognate mRNA, relative to its ability to inhibit its translation, the same enzyme preparation was active as translational repressor at a considerably lower protein/mRNA ratio, suggesting the two phenomena to be disconnected. Of interest is the fact that the presence of the enzyme protein N-terminal methionine proved to be critical for binding, but not for repression of its translation, indicating that mRNA binding requires a methionine or an adduct (i.e. methionine-histidine) at the N-terminus of TS, but that the translational repression effect does not. Notably, chicken liver dihydrofolate reductase, which is incapable of binding to T. spiralis TS mRNA, repressed the translation of TS.
旋毛虫是一种引起旋毛虫病的寄生线虫,其胸苷酸合成酶(TS)被发现能结合自身mRNA并抑制后者的翻译,这与其人类对应物的情况相似[朱、科勒、凯西、德雷克、查布纳、埃尔伍德、津恩和阿莱格拉(1991年)《美国国家科学院院刊》88,8977 - 8981]。然而,与人类TS形成鲜明对比的是,寄生虫酶与mRNA的相互作用不受任何底物(脱氧尿苷酸或N(5,10)-亚甲基四氢叶酸)的影响,也不受抑制剂(氟脱氧尿苷酸;单独使用或在N(5,10)-亚甲基四氢叶酸存在下)的影响——这与恶性疟原虫双功能酶的情况相似[张和拉托德(2002年)《科学》296,545 - 547]。此外,其他人(朱等人,1991年)发现能阻止人类TS抑制其翻译的相同配体,增强了寄生虫酶翻译的抑制作用。在比较TS结合其同源mRNA的能力与其抑制翻译的能力时,相同的酶制剂在蛋白质/ mRNA比例相当低时作为翻译阻遏物具有活性,这表明这两种现象没有关联。有趣的是,酶蛋白N端甲硫氨酸的存在被证明对结合至关重要,但对其翻译的抑制并非如此,这表明mRNA结合需要TS N端的甲硫氨酸或加合物(即甲硫氨酸 - 组氨酸),但翻译抑制作用则不需要。值得注意的是,鸡肝二氢叶酸还原酶不能结合旋毛虫TS mRNA,但能抑制TS的翻译。