Collaborative Research: Revealing essential regulatory proteins in tardigrade cryptobiosis

水熊虫（水熊）是八条腿的微小无脊椎动物，以其在极端压力下生存的能力而闻名。这种生存的标志是他们独特的能力，形成一个'tun'，一个生存状态，通过收回四肢，排出内部水储存，并显着降低新陈代谢实现。水熊虫可以保持这种状态多年，同时保持相对完整，只有当外部威胁被消除时才会出现。然而，这种存活是如何调节的在很大程度上是未知的。研究人员的工作揭示了水熊虫的生存依赖于高活性含氧化学物质的存在，这些化学物质是所有生命系统中存在的小细胞信使。这些化学物质是重要的信号分子，通过修饰细胞内的蛋白质来改变代谢活性。 来自查佩尔山的北卡罗来纳州大学和马歇尔大学的多学科科学家团队可以通过强大的分析和生物化学方法跟踪这些信号和受影响的生物化合物。研究小组将联合收割机结合这些方法，全面绘制水熊虫在独特压力下进入和离开隧道的入口和出现，使他们能够确定实现极端压力耐受性的精确适应。这项工作对于理解细胞内极端压力耐受性的分子策略是不可或缺的，这些策略可以应用于理解地球上生命的压力。 合作团队将在他们的校园创建姐妹课程，跨机构工作，以表征不同的缓步动物蛋白质。 此外，学生将从事本科研究，并获得蛋白质化学，计算建模和科学素养方面的经验。水熊虫（水熊虫）是世界性的微观无脊椎动物，使用巧妙的极端耐受模式对环境压力源做出快速反应，统称为隐生。了解管理缓步隐生的监管过程是必不可少的，以揭示暴露于极端压力时，保护生化途径的分子策略。虽然隐生现象在分类群中普遍存在，但对分子机制以及不同类型相互关联的程度只有初步的了解。我们还远远没有全面了解生物化学的参与者，不同网络之间的协调，或者压力和生存之间的相互作用。一个多学科小组将联合收割机结合他们在蛋白质组学和活性氧监测方面的专业知识，旨在阐明缓步动物极端耐受性的氧化还原依赖性。他们将在生理学和蛋白质组学水平上表征缓步动物的生存，从而增强对缓步动物在各种恶劣环境中生存的分子机制的理解。项目成果包括：a）鉴定和表征隐生菌诱导所需的基本氧化还原修饰蛋白; B）绘制不同隐生菌的氧化还原信号;和c）鉴定缓步动物“隐生菌组”中的关键调控点。这项工作将产生最全面的生物分子框架的隐生，到目前为止。这一奖项反映了NSF的法定使命，并已被认为是值得通过评估使用基金会的智力价值和更广泛的影响审查标准的支持。

Tardigrades (water bears) are eight-legged, microscopic invertebrates renowned for their ability to survive extreme stress. The hallmark of this survival is their unique ability to form a ‘tun,’ a survival state achieved through withdrawing of limbs, expelling internal water stores, and significantly decreasing metabolism. Tardigrades can remain in this state for years while remaining relatively undamaged, emerging only when the external threat has been removed. However, how this survival is regulated is largely unknown. The investigators' work has revealed a dependence of tardigrade survival on the presence of highly reactive oxygen-containing chemicals, small cellular messengers present in all living systems. These chemicals are essential signaling molecules that alter metabolic activity through the modification of proteins within the cell. The multidisciplinary team of scientists from the University of North Carolina at Chapel Hill and Marshall University can track these signals and affected biological compounds within tardigrades through powerful analytical and biochemical methods. The investigator team will combine these approaches to comprehensively map the entrance and emergence of tardigrade into and from tun across unique stresses allowing them to determine the precise adaptations that enable extreme stress tolerance. This work is integral to understanding molecular strategies for extreme stress tolerance within cells that can be applied to understand stress across life on earth. The collaborative teams will create sister courses on their campus, working across institutions to characterize different tardigrade proteins. In addition, students will be engaged in undergraduate research and gain experience in protein chemistry, computational modeling, and scientific literacy.Tardigrades (water bears) are cosmopolitan microscopic invertebrates that respond quickly to environmental stressors using ingenious modes of extremotolerance collectively known as cryptobiosis. Understanding the regulatory processes governing tardigrade cryptobiosis is essential to revealing the molecular strategies that preserve biochemical pathways when exposed to extreme stress. While cryptobiosis is prevalent across taxa, there exists only a nascent understanding of the molecular mechanisms and to what extent different types are interconnected. We are far from a comprehensive understanding of the biochemical participants, the coordination among diverse networks, or of the interplay between stress and survival. A multi-disciplinary team will combine their expertise in proteomics and reactive oxygen species monitoring with the aim of elucidating the redox-dependence of tardigrade extremotolerance. They will characterize tardigrade survival on both a physiological and proteomic level, enabling enhanced understanding of the molecular mechanisms through which tardigrades survive various hostile environments. Project outcomes include: a) the identification and characterization of essential redox-modified proteins required for cryptobiosis induction; b) the mapping of redox signaling across distinct cryptobioses; and c) the identification of key regulatory points in the tardigrade ‘cryptobiome’. This work will generate the most comprehensive biomolecular framework for cryptobiosis, to date.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.