Abstract：
Kinetoplastids, a group of flagellated protists that are often insect intestinal parasites, encounter various sources of oxidative stress. Such stressors include reactive oxygen species, both internally produced within the protist, and induced externally by host immune responses. This investigation focuses on the role of a highly conserved aspartate-based protein phosphatase, PTP-Interacting protein (PIP39) in managing oxidative stress. In addition to its well accepted role in a Trypanosoma brucei life stage transition, there is evidence of PIP39 participation in the T. brucei oxidative stress response. To examine whether this latter PIP39 role may exist more broadly, we aimed to elucidate PIP39\'s contribution to redox homeostasis in the monoxenous parasite Leptomonas seymouri. Utilizing CRISPR-Cas9-mediated elimination of PIP39 in conjunction with oxidative stress assays, we demonstrate that PIP39 is required for cellular tolerance to oxidative stress in L. seymouri, positing it as a putative regulatory node for adaptive stress responses. We propose that future analysis of L. seymouri PIP39 enzymatic activity, regulation, and potential localization to a specialized organelle termed a glycosome will contribute to a deeper understanding of the molecular mechanisms by which protozoan parasites adapt to oxidative environments. Our study also demonstrates success at using gene editing tools developed for Leishmania for the related L. seymouri.
摘要：
肌体，一群鞭毛的原生生物，通常是昆虫肠道寄生虫，遇到各种来源的氧化应激。这些应激源包括活性氧，都是在原生内部生产的，并由宿主免疫反应在外部诱导。这项研究的重点是高度保守的天冬氨酸为基础的蛋白磷酸酶的作用，PTP相互作用蛋白（PIP39）在氧化应激管理中的作用。除了在布鲁氏锥虫生命阶段过渡中被广泛接受的作用外，有证据表明PIP39参与了布鲁氏菌的氧化应激反应。为了检查后一种PIP39的作用是否可能更广泛地存在，我们的目的是阐明PIP39对单氧性寄生虫轻浮菌氧化还原稳态的贡献。利用CRISPR-Cas9介导的PIP39消除结合氧化应激测定，我们证明了PIP39是细胞对L.seymouri氧化应激的耐受性所必需的，将其定位为自适应应激反应的推定调节节点。我们建议未来分析L.seymouriPIP39酶活性，regulation,和潜在的定位到一个专门的细胞器称为糖体将有助于更深入地理解的分子机制，原生动物寄生虫适应氧化环境。我们的研究还证明了使用利什曼原虫开发的基因编辑工具对相关L.seymouri的成功。
