Collaborative Research: Revealing essential regulatory proteins in tardigrade cryptobiosis

缓步动物（水熊）是八足的微型无脊椎动物，以其在极端压力下生存的能力而闻名。这种生存的标志是它们形成“转”的独特能力，这是一种通过收回四肢、排出体内水分和显著降低新陈代谢来实现的生存状态。缓步动物可以在这种状态下保持数年，同时保持相对完好的状态，只有当外部威胁被消除时才会出现。然而，这种生存是如何调控的在很大程度上是未知的。研究人员的工作揭示了缓步动物的生存依赖于高活性含氧化学物质的存在，这些化学物质是存在于所有生命系统中的小细胞信使。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动。来自北卡罗来纳大学教堂山分校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化方法跟踪这些信号，并影响缓步动物体内的生物化合物。研究小组将结合这些方法，全面绘制缓步动物在独特压力下的进入和出现，使他们能够确定能够承受极端压力的精确适应。这项工作对于理解细胞内极端压力耐受的分子策略是不可或缺的，可以应用于理解地球上所有生命的压力。合作团队将在他们的校园里开设姐妹课程，跨机构合作，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。缓步动物（水熊）是一种普遍存在的微观无脊椎动物，它们对环境压力的反应非常迅速，它们采用了独特的极端耐受性模式，统称为隐生。了解缓步动物隐生的调控过程对于揭示在极端压力下保持生化途径的分子策略至关重要。虽然隐生现象在不同的分类群中普遍存在，但对分子机制和不同类型之间的相互联系程度只有初步的了解。我们对生物化学参与者，不同网络之间的协调，或压力与生存之间的相互作用还远远没有全面的了解。一个多学科团队将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。他们将在生理和蛋白质组学水平上描述缓步动物的生存特征，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)鉴定和表征诱导隐生所需的氧化还原修饰蛋白；B)不同隐生植物中氧化还原信号的映射；c)缓步动物“隐生群落”关键调控点的确定。这项工作将产生迄今为止最全面的隐生生物分子框架。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.