ROLE OF PFHO-1 IN P. FALCIPARUM INTRAERYTHROCYTIC DEVELOPMENT

PFHO-1 在恶性疟原虫红细胞内发育中的作用

• 批准号: 8662416
• 负责人: Daniel E. Goldberg
• 金额: $ 17.94万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-06 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8662416
• 关键词: Aminolevulinic Acid; Antimalarials; Binding; Cell Nucleus; Cells; ChIP-seq; Chloroquine; Country; DNA; DNA Binding; Development; Disease; Dominant-Negative Mutation; Erythrocytes; Escherichia coli; Food; Gene Deletion; Gene Expression; Gene Targeting; Genes; Genetic Engineering; Goals; Growth and Development function; Heme; Hemoglobin; Hemopexin; Homeostasis; Human; Knock-out; Knowledge; Malaria; Organism; Oxygenases; Parasites; Peptide Hydrolases; Peptides; Phenotype; Plasmodium falciparum; Play; Porphyrins; Production; Protease Inhibitor; Protein Binding; Protein Biosynthesis; Proteins; Reporter; Ribosomal RNA; Role; Signal Transduction; Source; Supplementation; System; Testing; Time; Transfer RNA; Vacuole; Viral; chemotherapy; heme a; heme biosynthesis; infancy; knockout gene; mutant; new technology; porphyrin a; protoporphyrin IX; public health relevance; research study; response

Abstract Exploration of the control of Plasmodium falciparum intraerythrocytic growth and development is in its infancy. Understanding the signals and effectors of parasite progression will enhance our knowledge of how this parasite survives in its host cell and will uncover new targets for chemotherapy. PfHO-1 is a heme oxygenase-like protein of unknown function expressed in intraerythrocytic malaria parasites. We have found that PfHO-1 has a porphyrin-responsive DNA- binding activity and associates preferentially with genes that are involved in protein synthesis. Our hypothesis is that PfHO-1 responds to heme accumulation from hemoglobin degradation and is crucial for intra-erythrocytic P. falciparum development. The goal of the study proposed here is to test this hypothesis. We will assess the response of PfHO-1 to perturbation of intracellular porphyrin levels in intraerythrocytic P. falciparum and in an E. coli reporter system. We will impair PfHO-1 action by a new double-positive selection gene knockout approach and by conditional expression of a porphyrin-unresponsive version of the protein. These studies will allow us to establish whether PfHO-1 plays an important role in sensing cellular porphyrin levels and participating in critical aspects of cellular homeostasis. The proposed experiments will also develop new technologies for porphyrin manipulation and for selection of deleterious phenotypes in reverse genetically engineered parasites.

抽象的 控制恶性疟原虫红细胞内生长和控制的探索 发展尚处于起步阶段。了解寄生虫的信号和效应器 进展将增强我们对这种寄生虫如何在宿主中生存的了解 细胞，并将发现化疗的新靶点。 PfHO-1 是一种血红素 红细胞内表达的功能未知的加氧酶样蛋白 疟疾寄生虫。我们发现 PfHO-1 具有卟啉响应 DNA- 结合活性并优先与参与的基因结合 蛋白质合成。我们的假设是 PfHO-1 对血红素积累作出反应 避免血红蛋白降解，对于红细胞内恶性疟原虫至关重要 发展。这里提出的研究的目的是检验这一假设。我们 将评估 PfHO-1 对细胞内卟啉扰动的反应 红细胞内恶性疟原虫和大肠杆菌报告系统中的水平。我们将 通过新的双阳性选择基因敲除损害 PfHO-1 的作用 方法并通过卟啉无反应版本的条件表达 蛋白质。这些研究将使我们能够确定 PfHO-1 是否发挥作用 在感知细胞卟啉水平和参与关键过程中发挥重要作用 细胞稳态的各个方面。拟议的实验还将发展 卟啉操作和有害物质选择的新技术 反向基因工程寄生虫的表型。