Department of Biology, Indiana University, Bloomington, Indiana, USA.
PLoS Pathog. 2010 Feb 5;6(2):e1000754. doi: 10.1371/journal.ppat.1000754.
Toxoplasma gondii is a leading cause of congenital birth defects, as well as a cause for ocular and neurological diseases in humans. Its cytoskeleton is essential for parasite replication and invasion and contains many unique structures that are potential drug targets. Therefore, the biogenesis of the cytoskeletal structure of T. gondii is not only important for its pathogenesis, but also of interest to cell biology in general. Previously, we and others identified a new T. gondii cytoskeletal protein, TgMORN1, which is recruited to the basal complex at the very beginning of daughter formation. However, its function remained largely unknown. In this study, we generated a knock-out mutant of TgMORN1 (DeltaTgMORN1) using a Cre-LoxP based approach. We found that the structure of the basal complex was grossly affected in DeltaTgMORN1 parasites, which also displayed defects in cytokinesis. Moreover, DeltaTgMORN1 parasites showed significant growth impairment in vitro, and this translated into greatly attenuated virulence in mice. Therefore, our results demonstrate that TgMORN1 is required for maintaining the structural integrity of the parasite posterior end, and provide direct evidence that cytoskeleton integrity is essential for parasite virulence and pathogenesis.
刚地弓形虫是导致先天性出生缺陷的主要原因,也是人类眼部和神经疾病的病因之一。其细胞骨架对于寄生虫的复制和入侵至关重要,并且包含许多潜在的药物靶点。因此,刚地弓形虫细胞骨架结构的生物发生不仅对其发病机制很重要,而且对一般的细胞生物学也很重要。以前,我们和其他人鉴定了一种新的刚地弓形虫细胞骨架蛋白 TgMORN1,它在子体形成的最初阶段被招募到基底复合物。然而,它的功能在很大程度上仍然未知。在这项研究中,我们使用 Cre-LoxP 系统生成了 TgMORN1 的敲除突变体(DeltaTgMORN1)。我们发现 DeltaTgMORN1 寄生虫中的基底复合物的结构受到严重影响,胞质分裂也存在缺陷。此外,DeltaTgMORN1 寄生虫在体外的生长受到严重损害,这导致其在小鼠体内的毒力大大减弱。因此,我们的结果表明 TgMORN1 对于维持寄生虫后端的结构完整性是必需的,并提供了直接证据表明细胞骨架完整性对于寄生虫的毒力和发病机制是必不可少的。