Identification and analysis of novel tardigrade stress response mechanisms

摘要 细胞稳定性对于在极端环境中生存至关重要。一些细胞和生物体可以在 即使在其组成部分不稳定的环境中，这表明细胞保护剂必须 在这种环境下生存的能力。这种保护剂具有用于生物医学的潜力。 应用，如稳定敏感的生物材料，以及它们发挥作用的机制， 基本的生物学兴趣。本提案的目标1旨在确定缓步动物中的细胞保护剂。 缓步动物是无脊椎动物，可以承受各种各样的环境压力，包括 干燥、冰冻、辐射、高压，甚至太空的真空。以前的工作已经确定了一个 有限数量的缓步动物保护蛋白，并且值得注意的是，这些蛋白质可以赋予对 在细菌和酵母中脱水，在人类细胞中增强辐射耐受性。这些研究奠定了 为筛选新型缓步动物应激保护蛋白奠定基础。由此确定的利益保护者 筛选也将在人细胞培养物中表达，以研究细胞稳定性的机制。 建立，并确定蛋白质的强有力的候选人，以提高生物样品的稳定性。的 该建议的第二个目的是揭示滞育的遗传调控--滞育是一种发育暂停的状态 并增强了对压力的抵抗力。控制滞育的途径也可能调节 生产压力保护剂。因此，鉴定保守的细胞信号传导途径所需的 滞育将为未来寻找保护剂的研究提供信息。这项工作完成后， 新的保护剂分子和机制的压力抵抗，可能证明有用的应用， 生物材料的储存。

Abstract Cellular stability is essential for surviving extreme environments. Some cells and organisms can survive in environments even in which their constituent parts are not stable, suggesting that cellular protectants must exist that allow for survival in these contexts. Such protectants have the potential to be utilized for biomedical applications like stabilizing sensitive biological materials, and the mechanisms by which they function are of fundamental biological interest. Aim 1 of this proposal seeks to identify cellular protectants in tardigrades. Tardigrades are invertebrates that can withstand a remarkable variety of environmental stresses including desiccation, freezing, radiation, high pressure, and even the vacuum of space. Previous work has identified a limited number of tardigrade protectant proteins, and remarkably these proteins can confer resistance to desiccation in bacteria and yeast and enhanced radiotolerance in human cells. These studies lay the groundwork to screen for novel tardigrade stress protectant proteins. Protectants of interest identified from this screen will also be expressed in human cell culture in order to study mechanisms by which cellular stability is established, and to identify proteins that are strong candidates for improving stability of biological samples. The second aim of this proposal is to uncover genetic regulation of diapause – a state of suspended development and increased stress resistance – in tardigrades. The pathways that control diapause may also regulate production of stress protectants. Therefore, identification of conserved cell signaling pathways required for diapause will inform future studies searching for protectants. Completion of this work will lead to identification of novel protectant molecules and mechanisms of stress resistance that may prove useful for applications like storage of biomaterials.