Determinants of apicoplast maintenance in malaria parasites

疟疾寄生虫含有一种名为顶体的叶绿体细胞器，它是寄生虫在 用于人类和传播给蚊子。顶生质体一直被认为是一种重要的来源 新的药物靶点来对抗不可避免的抗药性问题，然而，事实证明很难 鉴定和验证对寄生虫生存至关重要的质外体蛋白。这个目标现在是可以实现的。 使用新的遗传工具结合新陈代谢旁路的顶生质体。治疗血液期寄生虫 与异戊二烯前体IPP(异戊烯基焦磷酸)一起，质外体抑制物存活下来-即使是那些 导致细胞器的破坏和细胞器基因组的丢失。我们在这一发现的基础上创建了一个 新陈代谢旁路寄生菌株系，在胞浆中产生类异戊二烯前体，使用基因工程的四- 甲氧戊酸酶途径。我们建议结合使用遗传方法和 新陈代谢旁路，以确定所有核编码的蛋白质，这些蛋白质对质外体功能和 寄生虫的生存。我们还将使用新的条件工具(击倒、条件本地化、DiCre)来进一步 描述特定蛋白质的作用和与其丢失相关的表型。我们的实验将 帮助建立更完整的代谢途径和所需的非代谢过程的图景 质外体功能与寄生虫存活。最终，我们打算确定新的目标，并验证已知的 未来治疗疟疾和阻止其传播的药物开发的目标。

Malaria parasites contain a plastid organelle called the apicoplast that is required for parasite survival in humans and for transmission to mosquitoes. The apicoplast has long been recognized as an important source of new drug targets to combat the inevitable problem of drug resistance, however, it has proven difficult to identify and validate apicoplast proteins that are essential for parasite survival. This goal is now achievable using new genetic tools in combination with metabolic bypass of the apicoplast. Blood stage parasites treated with the isoprenoid precursor IPP (isopentenyl pyrophosphate) survive apicoplast inhibitors - even those which result in disruption of the organelle and loss of the organellar genome. We built on this finding by creating a metabolic bypass parasite line that produces isoprenoid precursors in the cytosol using an engineered four- enzyme mevalonate pathway. We propose to use a combination of genetic approaches in conjunction with metabolic bypass to identify all nuclear-encoded proteins which are essential for apicoplast function and parasite survival. We will also use new conditional tools (knockdown, conditional localization, DiCre) to further characterize the roles of specific proteins and the phenotypes associated with their loss. Our experiments will help to build a more complete picture of the metabolic pathways and non-metabolic processes required for apicoplast function and parasite survival. Ultimately, we intend to identify novel targets and to validate known targets for future development of drugs to cure malaria and stop its transmission.