Molecular analysis of a kinase essential for replication of Plasmodium falciparum

恶性疟原虫复制所必需的激酶的分子分析

基本信息

批准号：
7868632
负责人：
JEFFREY D DVORIN
金额：
$ 13.23万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2013-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7868632
关键词：
Advisory Committees Affinity Age-Years Antimalarials Biochemical Biological Biological Process Biology Blood Boston Calcium Candidate Disease Gene Cells Cessation of life Child Mortality Clinical Commit Data Development Plans Drug Delivery Systems Drug resistance Environment Enzymes Epitopes Erythrocytes Evaluation Faculty Family Future Genes Genetic Goals HIV Human Human Genome Immunoelectron Microscopy Immunofluorescence Immunologic In Vitro Incubated Infection Knock-out Life Cycle Stages Malaria Mass Spectrum Analysis Mediating Mentorship Molecular Molecular Analysis Molecular Biology Palmitic Acylation Site Parasites Parasitology Pathogenesis Pathway interactions Pattern Pediatric Hospitals Peptide Hydrolases Pharmaceutical Preparations Phosphorylation Phosphotransferases Plants Plasmodium falciparum Positioning Attribute Process Proteins Proteomics Public Health Schools Recombinants Regulation Reporting Research Resistance Resistance development Role Scanning Signal Transduction Staging System Techniques Testing Text Therapeutic Time Training Transgenic Organisms Tropical Medicine Vaccines Work abstracting asexual base calcium-dependent protein kinase career career development design enzyme activity experience genetic manipulation in vivo novel pathogen preference prevent programs protein transport research study trafficking

项目摘要

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.

描述（由申请人提供）：注意：此摘要是从申请中提取的，未经 SRA 验证。当申请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，其中大部分死亡是由恶性疟原虫引起的。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。在寄生虫生命周期的无性复制阶段，恶性疟原虫需要有效地侵入和离开人类红细胞。我们的初步数据已经确定了一种类植物钙依赖性蛋白激酶 PfCDPK5，它对于恶性疟原虫的排出至关重要。这些实验首次实现了恶性疟原虫中重要血液阶段基因的功能性敲除。 PfCDPK5 缺陷的寄生虫在红细胞内生命周期的最后阶段停滞。我们假设 PfCDPK5 介导恶性疟原虫排出所需的关键钙依赖性信号。该提案的目标是更好地了解 PfCDPK5 功能并阐明其在寄生虫从受感染的红细胞中排出的过程中的作用。第一个目标是，候选人将确定这种重要激酶的定位、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，第二个目标的实验将鉴定 PfCDPK5 的体内底物。这些研究将在恶性疟原虫遗传学和耐药机制领域的先驱 Dyann Wirth 博士的指导下进行，并由恶性疟原虫基因操作和分子生物学专家 Manoj Duraiseh 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以及接受他的科学顾问委员会的指导和指导来实现。该候选人的长期职业目标是获得终身教职并继续他对疟疾分子发病机制的研究。哈佛大学公共卫生学院的培训机会以及博士的指导。 Wirth 和 Duraiseh 是该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，并保证候选人能够将至少 75% 的全职精力投入到本提案中描述的活动中。（相关性）：疟疾是全球五岁以下儿童死亡的第四大原因，其中大部分死亡是由恶性疟原虫感染引起的。该应用的目的是更好地了解恶性疟原虫在人类红细胞中复制的分子机制。该项目将利用遗传和生化技术，描述寄生虫从红细胞中排出的关键生物过程，并最终确定未来抗疟药物的新靶点。