Molecular analysis of a kinase essential for replication of Plasmodium falciparum

描述（由申请人提供）： 注意事项：本摘要摘自应用程序，未经SRA验证。当应用程序扫描过程出现问题时，提取的文本可能不正确或不完整。 ================== 疟疾是全球五岁以下儿童死亡的第四大原因，其中大多数是恶性疟原虫造成的。对恶性疟原虫生命周期的分子理解将有助于新疗法的合理设计。恶性疟原虫需要在寄生虫生命周期的无性复制阶段有效地侵入和排出人红细胞。我们的初步数据已经确定了植物样钙依赖性蛋白激酶PfCDPK5，这是恶性疟原虫出口的关键。这些实验产生了恶性疟原虫中一个重要血液阶段基因的第一个功能性敲除。PfCDPK5缺陷型寄生虫在红细胞内生命周期的非常晚期停止。我们假设PfCDPK5介导恶性疟原虫逃逸所需的关键钙依赖性信号。该提案的目标是更好地了解PfCDPK5的功能，并阐明其在寄生虫从受感染的红细胞中排出中的作用。在第一个目标中，候选人将确定这种必需激酶的定位，运输和调节。使用两种基于质谱的技术和第三种候选基因方法，第二个目的的实验将鉴定PfCDPK5的体内底物。这些研究将在恶性疟原虫遗传学和耐药性机制的先驱Dyann Wirth博士的指导下进行，并与恶性疟原虫遗传操作和分子生物学专家Manoj Duraisingh博士共同指导。除了拟议的实验外，候选人的职业发展目标是获得寄生虫学，恶性疟原虫分子生物学和热带医学方面的专业知识。这些近期目标将通过参加蛋白质组学、寄生虫生物学和热带医学的正式课程，并接受他的科学咨询委员会的指导和指导来实现。候选人的长期职业目标是获得终身教职，并继续研究疟疾的分子发病机制。哈佛公共卫生学院的培训机会以及Wirth和Duraisingh博士的指导是这个职业发展计划的理想环境。波士顿儿童医院致力于这一职业发展计划，并已保证候选人将能够投入至少75%的全职努力，在本建议书中描述的活动。（相关性）：疟疾是全球五岁以下儿童死亡的第四大原因，其中大多数死亡是由恶性疟原虫感染造成的。本申请的目的是更好地了解恶性疟原虫在人红细胞中复制的分子机制。利用遗传和生化技术，该项目将表征寄生虫从红细胞中逸出的关键生物过程，并最终确定未来抗疟疾药物的新靶点。

DESCRIPTION (provided by applicant): NOTICE: THIS ABSTRACT WAS EXTRACTED FROM APPLICATION AND HAS NOT BEEN PROOFED BY AN SRA.WHEN THERE ARE PROBLEMS WITH THE APPLICATION SCANNING PROCESS, THE EXTRACTED TEXT MAY BE INCORRECT OR INCOMPLETE. ================== Malaria is the fourth leading cause of mortality for children under five years of age globally, with most of these deaths resulting from Plasmodium falciparum. A molecular understanding of the life cycle of P. falciparum will facilitate the rational design of new therapies. P. falciparum requires efficient invasion into and egress out of human erythrocytes during the asexual replication stage of the parasite life cycle. Our preliminary data have identified a plant-like calcium-dependent protein kinase PfCDPK5 that is crucial for P. falciparum egress. These experiments have generated the first functional knockout of an essential blood-stage gene in P. falciparum. PfCDPK5-deficient parasites arrest at a very late-stage of the intra-erythrocytic life cycle. We hypothesize that PfCDPK5 mediates a critical calcium-dependent signal required for P. falciparum egress. The goals of this proposal are to gain a better understanding of PfCDPK5 function and elucidate its role in parasite egress from infected erythrocytes. In the first aim, the candidate will determine the localization, trafficking, and regulation of this essential kinase. Using two mass spectrometry based techniques and a third candidate gene approach, the experiments of the second aim will identify the in vivo substrate(s) of PfCDPK5. These studies will be conducted under the mentorship of Dr. Dyann Wirth, a pioneer in P. falciparum genetics and mechanisms of drug resistance, and the co-mentorship of Dr. Manoj Duraisingh, an expert in P. falciparum genetic manipulation and molecular biology. In addition to the proposed experiments, the candidate's career development goals are to gain expertise in parasitology, P. falciparum molecular biology, and tropical medicine. These immediate goals will be achieved by attending formal courses in proteomics, parasite biology, and tropical medicine and by receiving mentorship and guidance from his scientific advisory committee. The candidate's long-term career goals are to attain a tenure-track faculty position and to continue his research on the molecular pathogenesis of malaria. The training opportunities at the Harvard School of Public Health together with the mentorship of Drs. Wirth and Duraisingh are an ideal environment for this career development program. Children's Hospital Boston is committed to this career development plan and has assured that the candidate will be able to devote at least 75% full-time effort to the activities described in this proposal. (Relevance): Malaria is the fourth leading cause of mortality for children under five years of age globally, with most of these deaths resulting from Plasmodium falciparum infection. The goal of this application is gain a better understanding of the molecular mechanisms of P. falciparum replication in human red blood cells. Using genetic and biochemical techniques, this project will characterize the key biological process of parasite egress from red blood cells and ultimately identify new targets for future anti- malarial medications.