Role of Translesional Polymerases in Genome Diversification of the Malaria Parasite

项目摘要/摘要： 疟疾仍然是热带和热带地区居民发病和死亡的主要原因。 世界上的亚热带地区。寄生虫持续产生序列多样性的能力 在他们的基因组中是不能形成有效的红血球阶段的主要因素 疫苗和一直存在的抗药性问题。然而，我们的主要差距仍然存在 了解寄生虫基因组是如何维持的。我们的长期目标是找出 对维持寄生虫基因组和了解这些修复途径如何影响 可变性和基因组可塑性。 跨损伤(TLS)聚合酶是一种特殊的DNA聚合酶，能够继续DNA 通过损坏碱基和困难的模板进行合成，尽管它们在容易出错的情况下完成了这一点 举止。在模型系统中，TLS聚合酶产生了大多数新的突变。这里只有 灵长类疟疾基因组中存在两个TLS聚合酶，REV1和POLζ。我们的假设是 这些聚合酶在寄生虫基因组的维持中起着至关重要的作用，并有助于 和疟疾的发病机制：a)通过促进抗原多样性的产生 多拷贝半同源但不完全相同成员间的同源重组 基因家族和b)在整个基因组和在 转向抗药性的发展。使用基因组编辑技术、突变和SMRT 通过测序技术，我们试图揭示TLS聚合酶在人类疟疾中的作用 寄生虫，恶性疟原虫。 我们提议的研究将揭示疟原虫DNA修复的独特方面，这些方面将对 了解疟疾和那些研究基因组维护的人。我们的目标是精心设计的 对疟疾的重要临床方面有直接影响，寄生虫的发展倾向 抗药性和逃避宿主免疫系统。这项研究解决了一个重要而被忽视的问题 寄生虫生物学的一个方面。

Project Summary/Abstract: Malaria continues to be a major cause of morbidity and mortality among people living in the tropical and subtropical regions of the world. The ability of parasites to continuously generate sequence diversity within their genomes is a major contributor to the inability to develop effective erythrocytic stage vaccines and to the ever-present problem of drug resistance. Yet, key gaps remain in our understanding of how the parasite genome is maintained. Our long-term goal is to identify pathways that are crucial to maintaining the parasite genome and understand how these repair pathways impact mutability and genome plasticity. Translesion (TLS) polymerases are specialized DNA polymerases that are capable of continuing DNA synthesis through damage bases and difficult templates, though they accomplish this in an error prone manner. In model systems, TLS polymerases generate the majority of novel mutations. There are only two TLS polymerase present in the primate malaria genomes, Rev 1 and pol ζ. Our hypothesis is that these polymerases play a crucial role in parasite genome maintenance and contribute to persistence of and pathogenesis of malaria by a) driving the generation of antigenic diversity by promoting homologous recombination (HR) between semi-homologous but non-identical members of multicopy gene families and b) contributing to the generation of sequence variation throughout the genome and in turn to the development of drug resistance. Using genome editing techniques, mutagenesis and SMRT sequencing techniques, we seek to uncover the role of TLS polymerases in the human malaria parasite, Plasmodium falciparum. Our proposed research will uncover unique aspects of Plasmodium DNA repair that will be important for understanding malaria and to those that study genome maintenance in general. Our aims are designed to have direct impact on the important clinical aspects of malaria, the parasite's propensity to develop drug resistance and evade the host immune system. This study addresses an important and neglected aspect of parasite biology.