TRYPANOSOME REGULATORY PROTEIN

布氏毛滴虫VSG表达位点包含多个表达位点 在特定阶段高水平表达的相关基因 举止。我们之前已经克隆并测序了一个ESAG，我们将其命名为T- LR(代表富含亮氨酸的锥虫重复序列)和其他人称之为ESAG 8。 从T-LR预测的蛋白质序列包含两个有趣的结构域。 一种与调控蛋白中的环指基序具有同源性 以序列特定的方式结合DNA，而另一种是富含亮氨酸的 重复序列(LRR)广泛存在于原核生物和真核生物中 参与蛋白质相互作用的蛋白质。因此，很可能 T-LR在寄生虫生理中起着重要的调节作用。整体而言 本项目的目的是阐明T-LR的作用，并确定 其功能背后的分子机制。我们还计划确定和 描述T-LR可能与之相互作用的其他调节分子。我们 将在细菌、酵母、昆虫或 哺乳动物表达系统并产生针对该病毒的特异性抗体 潜在的核酸和蛋白质结合域。抗体会 被用来表征T-LR蛋白的大小，后 翻译修饰，以及特定阶段的表达和确定 它的亚细胞位置。然后我们将识别相互作用的分子 与锥虫细胞内的T-LR结合。这些核酸和蛋白质 将被定性并克隆他们的基因。此外，我们还将 通过基因敲除调控T-LR基因的正常表达 T-LR基因在血液中的表达及T-LR的过度表达 血流和前循环的形式。锥虫生长的变化， 其他特定基因的形态和表达将识别细胞 涉及T-LR的进程。这些研究旨在找出新的 分子过程为新型材料的开发提供了基础 寄生虫控制策略。此外，与其他几项研究一样， 在布氏锥虫中的现象，它们也应该提供对 所有真核生物的基础分子生物学。

The VSG expression site of T. brucei contains several Expression Site Associated Genes which are expressed at high levels in a stage-specific manner. We have previously cloned and sequenced an ESAG which we named T- LR (for Trypanosome Leucine-rich Repeat) and others have called ESAG 8. The protein sequence predicted from T-LR contains two interesting domains. One has homology to the RING finger motif in regulatory proteins which bind DNA in a sequence-specific fashion, while the other is a leucine-rich repeat (LRR) sequence found in wide variety of prokaryotic and eukaryotic proteins involved in protein-protein interactions. Thus, it is likely that T-LR has an important regulatory role in parasite physiology. The overall aim of this project is to elucidate the role of T-LR and to identify the molecular mechanisms underlying its function. We also plan to identify and characterize other regulatory molecules with which T-LR may interact. We will express recombinant T-LR proteins in bacterial, yeast, insect or mammalian expression systems and produce specific antibodies against the potential nucleic acid- and protein-binding domains. The antibodies will be used to characterize the T-LR protein in terms of size, post- translational modification, and stage-specific expression and to determine its subcellular location. We will then identify molecules which interact with T-LR within the trypanosome cell. These nucleic acids and proteins will be characterized and their genes cloned. In addition, we will modulate the normal expression of the T-LR gene by knockout of the expressed T-LR gene in bloodstream forms and over-expression of T-LR in bloodstream and procyclic forms. Changes in trypanosome growth, morphology and expression of other specific genes will identify cellular processes involving T-LR. These studies are designed to identify new molecular processes that provide the basis for development of novel parasite control strategies. In addition, like the study of several other phenomena in T. brucei, they should also provide new insights into the fundamental molecular biology of all eukaryotic organisms.