Study on 14-3-3 related proteins and signal transduction for control of African tyrpanosomiasis.

蛋白质14-3-3是一种富含中枢神经系统的蛋白质，广泛存在于动植物等真核生物中。已报道的与14-3-3结合的蛋白质很多，估计该蛋白在细胞周期、细胞凋亡等信号转导途径中具有重要作用。2)尽管14-3-3的差异模体保守，但锥虫14-3-3的特性与哺乳动物有很大的不同。3)我们构建了唯一有效的载体pQuadra(PQ)，在诱导前进行dsRNAi，没有泄漏。4)在14-3-3应用PQ进行dsRNAi检测原环型(Pr)。在形态上，PR被破坏为奇怪的多鞭毛形式和定向的动粒或核传递。5)用PQ诱导dsRNAi后，血流形态细胞(Bs)的生长受到抑制。6)bS细胞的形态变化与蛋白磷酸(PP)抑制剂冈田酸的作用非常相似。根据14-3-3与PP的关系分析，我们非常接近非洲锥虫病的治疗目标。

Protein 14-3-3 was described to be rich in CNS and ubiquitously found in eukaryotic organisms of plants and animals. Quite a many proteins have been reported to bind to 14-3-3 then it is estimated the protein has important roles in the cell cycle, apoptosis and other signal transduction paths.We examined 14-3-3 of Trypanosoma b.brucei (Tb) and remark the followings.1) The genes of 2 isotypes (I, II) of 14-3-3 could be cloned. The result produced the recombinant proteins and sensitive mono-specific rat antibodies against them.2) Though 14-3-3 distinctive motifs are conserved, the characteristics of trypanosome 14-3-3 are quite different form those of mammals.3) We developed solely effective vector construct named pQuadra (pQ) to perform dsRNAi without leakage before induction.4) Procyclic forms (pr) were examined by dsRNAi on 14-3-3 applying pQ. Morphologically pr was damaged to bizarre multiflagellated form and disoriented kinetoplsts or nuclei delivery. The growth of pr was suppressed.5)On blood stream forms (bs) after induction of dsRNAi with pQ more drastic effect appeared earlier than pr and the growth was entirely suppressed6) The morphological change of bs was very similar to the effect of okadaic acid addition to culture of bs, which is protein phosphates (pp) inhibitor. Analysis based on relation between 14-3-3 and pp we came very close to the therapeutic target of African trypanosomiasis.