Unique functions of the mitochondrial tRNA import machinety in T. gondii

简介（由申请人提供）：申请人Lilach Sheiner在刚地弓形虫基因操作方面取得了很高的成就，并且最近开发了一个管道来识别和表征寄生虫内共生细胞器独特成分的编码基因。在指导阶段，申请人将利用该系统，专注于鉴定和破坏可能适合作为新药靶点的线粒体tRNA进口（MITI）的寄生虫特异性成分。此外，候选人将开发工具来表征MITI组件的生物功能，并将在导师的帮助下掌握相关的生化技能。作为基础生物医学研究机构的独立研究员，她将应用这些分子和生化工具来研究MITI和其他寄生虫线粒体生物发生的特定方面，作为药物靶点的有希望的来源。CTEGD培养了一个高度合作的研究环境，有19名教师对寄生虫生物学的各个方面感兴趣，并拥有优秀的核心设施。导师Boris Striepen是弓形虫内共生生物学方面的专家，也是弓形虫分子研究领域的领军科学家之一。共同导师Stephen Hajduk是一位生物化学家专家，在寄生虫线粒体生物发生方面有着广泛的经验，他对MITI在寄生虫方面有特殊的兴趣。弓形虫线粒体是必不可少的，并显示出几个独特的特征。最近的数据表明，这些寄生虫的MITI可能依赖于一种不同的机制。本提案的目标是利用一种新的生物信息学搜索策略，确定导致这种分化的成分，并通过反向遗传学解决它们的功能，使用一种新的流线型系统来产生条件突变体。虽然一些工具可以表征刚地弓形虫的tRNA输入和线粒体功能，但其他工具仍有待开发和适应。纯线粒体的分离尚未建立，这限制了生物化学研究的范围。这一挑战将在拟议的项目中得到解决。这些研究将为弓形虫线粒体的生物发生开辟新的视角，从而有助于设计新的对策，也与其他顶复合体寄生虫，如疟疾病原体恶性疟原虫有关。

DESCRIPTION (provided by applicant): The applicant, Lilach Sheiner, is highly accomplished in Toxoplasma gondii genetic manipulation, and has recently developed a pipeline to identify and characterize genes encoding unique components of parasite endosymbiotic organelles. During the mentored phase, the applicant will utilize this system and focus on the identification and disruption of parasite-specific components of mitochondrial tRNA import (MITI) that are likely suitable as new drug targets. Furthermore the candidate will develop tools to characterize the biological function of MITI components, and will master the relevant biochemical skills, with the help of the mentors. As an independent investigator at an institution of basic biomedical research, she will apply these molecular and biochemical tools to study MITI and other parasite-specific aspects of mitochondrial biogenesis, as a promising source for drug targets. The CTEGD fosters a highly collaborative research environment with 19 faculty members interested in various aspects of parasite biology and with excellent core facilities. The mentor, Boris Striepen, is an expert on endosymbiont biology in T. gondii and is one of the leading scientists in the field of molecular research in these parasites. The co-mentor, Stephen Hajduk, is an expert biochemist with wide ranging experience in parasite mitochondria biogenesis, and who has specific interest in MITI in parasites. The T. gondii mitochondrion is essential and shows several unique features. Recent data suggests that MITI in these parasites is likely to be dependent on a divergent machinery. The goal of this proposal is to identify the components responsible for this divergence, utilizing a novel bioinformatics search strategy, and to address their function by reverse genetics, using a new streamlined system to generate conditional mutants. While some tools to characterize tRNA import and mitochondrial function in T. gondii are at hand, others await development and adaptation. Isolation of pure mitochondria has not been established, which limits the extent of biochemical research available. This challenge will be addressed in the proposed project. These studies will open new perspectives on the biogenesis of the T. gondii mitochondrion and hence contribute to the design of new countermeasures, also of relevance to other apicomplexan parasites such as the causative agent of malaria, Plasmodium falciparum.