MOLECULAR ANALYSIS OF MALARIA MITOCHONDRIAL GENE REGULATION

人类疟疾寄生虫恶性疟原虫迅速进化出抗药性，创造了 迫切需要新的治疗策略。发现和开发新技术的关键障碍 药物开发目标是关于最基本的过程和监管的知识鸿沟 小路。理想情况下，新目标应该高度保守，不能或能力有限。 变异为了进化出抵抗力。寄生虫的线粒体功能至关重要 与人类细胞器有很大的不同；然而，大多数 尚未在疟疾寄生虫中鉴定出线粒体蛋白。在红细胞内注射期间 发展周期(IDC)，恶性疟原虫由一个含有大约 约20个拷贝的6kb基因组，以广泛重组为特征。IDC寄生虫 线粒体不会产生插入电子传递链(ETC)酶的隆起，因此 细胞器可能进化出独特的转录或翻译抑制系统来限制 线粒体编码基因的表达。来自我们的综合方法的结果 全基因组测序和代谢图谱表明线粒体基因之间存在联系 表达调控和对ETC抑制剂的抗性，可能是由于重组。因此， 我们假设1)多拷贝状态的一个特征是隐藏的线粒体的保留 基因组拷贝编码突变的等位基因，这些基因可以重组以求生存。2)恶性疟原虫 线粒体使用以前未描述的基因表达系统，用于抑制和 这种细胞器所特有的激活。目前的目标是找出 线粒体基因组重组及其在耐药中的作用 确定线粒体DNA修复途径和转录机制 线粒体使用单细胞器方法。该建议书将确定和定义以前 寄生虫生物学的未知和未描述的方面，目标是推动理性 药物设计。这些研究将提炼我们对基因基本机制的认识 疟疾线粒体的调控。研究支撑这些效应的机制 将导致高度保守的药物开发目标的确定。

The human malaria parasite, Plasmodium falciparum, rapidly evolves drug resistance, creating the urgent need for new treatment strategies. A critical barrier to identifying and developing new drug development targets is a knowledge gap regarding most essential processes and regulatory pathways. Ideally, new targets should be highly conserved and be unable or have limited ability to mutate in order to evolve resistance. Parasite mitochondrial function is critically essential across all the life stages and differs substantially from the human organelle; however, most mitochondrial proteins have yet to be identified in malaria parasites. During the intraerythrocytic development cycle (IDC), P. falciparum is supported by a single mitochondrion containing about ~20 copies of the 6 kb genome, characterized by extensive recombination. IDC parasite mitochondria do not make cristae to insert electron transport chain (ETC) enzymes, thus the organelle may have evolved unique transcription or translation repression systems to limit expression of the mitochondrial encoded genes. Results from our integrative approach combining whole genome sequencing and metabolic profiling, suggests a link between mitochondrial gene expression regulation and resistance to ETC inhibitors, potentially due to recombination. Thus, we hypothesize that 1) a feature of the multicopy status is retention of cryptic mitochondrial genome copies encoding mutant alleles, which can be recombined for survival. 2) P. falciparum mitochondria use previously uncharacterized gene expression systems, for repression and activation which are unique to this organelle. The current objectives are to identify the source of recombination between mitochondrial genomes and its contribution to drug resistance as well as to determine the mitochondrial DNA repair pathways and transcriptional machinery of the mitochondria using single-organelle approaches. This proposal will identify and define previously unknown and uncharacterized aspects of parasite biology, with the goal of advancing rational drug design. These studies will refine our knowledge about the basic mechanism of gene regulation in the malaria mitochondria. Investigating the mechanisms underpinning these effects will lead to the identification of highly conserved drug development targets.