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

蛋白14-3-3被描述为富含CNS并且普遍存在于植物和动物的真核生物中。14 -3-3是布氏锥虫（Trypanosoma B.brucei，Tb）的一种重要的蛋白质，它与14 - 3 -3结合，在细胞周期、细胞凋亡等信号转导途径中发挥重要作用。2）14-3-3序列虽然保守，但其结构域的保守性很强，14-3-3锥虫的特性与哺乳动物锥虫的特性有很大的不同。3）我们开发了唯一有效的载体pQuadra（pQ），用于在诱导前无渗漏地进行dsRNAi。4）对14-3- 3锥虫的前环形式（pr）进行了dsRNAi检测。3应用pQ.形态学上，pr被破坏成奇异的多鞭毛形式和定向的动质体或核递送。5）pQ诱导dsRNAi后，对血流形态（bs）的影响比pr更明显，且生长完全被抑制。6）bs形态变化与蛋白磷酸（pp）抑制剂冈田酸对bs的影响非常相似。根据14-3-3和pp之间的关系分析，我们非常接近非洲锥虫病的治疗靶点。

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.