Ouna Benard Aswani, Stewart Mhairi, Helbig Claudia, Clayton Christine
Zentrum fur Molekular Biologie Heidelberg, ZMBH-DKFZ Alliance, Im Neuenheimerfeld 282, Heidelberg 69120, Germany.
Mol Biochem Parasitol. 2012 Jun;183(2):184-8. doi: 10.1016/j.molbiopara.2012.02.006. Epub 2012 Feb 17.
CCCH-type zinc finger proteins have various roles in RNA metabolism. We here analysed the functional relevance of two such proteins from Trypanosoma brucei, TbZC3H12 and TbZC3H13. Each protein has a single CCCH motif very similar to those seen in metazoan proteins that regulate mRNA degradation. TbZC3H12 is expressed in bloodstream form parasites at low levels. It is phosphorylated, cytosolic and not required for normal growth of cultured bloodstream trypanosomes. RNA interference targeting TbZC3H13, on a TbZC3H12 null background, also had no effect on bloodstream trypanosome growth, but over-expression of tagged TbZC3H13 inhibited procyclic trypanosome growth. Tandem affinity purification of both proteins revealed various interesting potential interactions; specificity was assessed against a list of proteins that were found in 24 other pull-down experiments, which is provided. The conservation of TbZC3H12 in all kinetoplastids, and TbZC3H13 in Salivaria, suggests that the two proteins may be required for optimal growth at some stage of the parasite life-cycle.
CCCH型锌指蛋白在RNA代谢中具有多种作用。我们在此分析了来自布氏锥虫的两种此类蛋白TbZC3H12和TbZC3H13的功能相关性。每种蛋白都有一个单一的CCCH基序，与后生动物中调节mRNA降解的蛋白中的基序非常相似。TbZC3H12在血流形式的寄生虫中低水平表达。它被磷酸化，存在于细胞质中，对于培养的血流锥虫的正常生长不是必需的。在TbZC3H12基因缺失背景下，靶向TbZC3H13的RNA干扰对血流锥虫的生长也没有影响，但标记的TbZC3H13的过表达抑制了前循环锥虫的生长。对这两种蛋白进行串联亲和纯化揭示了各种有趣的潜在相互作用；针对在其他24次下拉实验中发现的一系列蛋白评估了特异性，这些蛋白列表已提供。TbZC3H12在所有动基体中保守，而TbZC3H13在唾液虫亚目中保守，这表明这两种蛋白可能是寄生虫生命周期某些阶段最佳生长所必需的。
