Animal disaggregases and amyloid based contributions to early animal development

酶法催化剂能够解开潜在的致病蛋白聚集体已被提出作为治疗神经退行性疾病的解决方案。考虑到所有其他形式的非动物生命都已经有这些酶，这就提出了一个问题：为什么人类失去了表达这些潜在有益疗法的能力？动物特异性蛋白质解缠酶缺失的答案可能在于蛋白质聚集体的潜在有益作用，这种作用与能够破坏它们的强效酶的存在是不相容的。在这项资助中，研究人员将利用两种强大的遗传模式生物的互补特征，以一种全面的方式测试这一假设。新发现的和潜在的更微妙的蛋白质聚集的酶调节将使用酵母模型系统进行鉴定，然后测试最近在线虫的年轻健康胚胎中发现的蛋白质聚集的调节。这笔拨款将对蛋白质聚集体及其专用酶调节因子进行全面检查，并试图了解它们如何作为控制早期动物发育关键事件的新机制，包括可以根据需要释放的关键发育蛋白质的储存。为了进一步加强科学与社会之间的联系，这笔资金还将用于发展新的举措，让中学生、教师和本科生了解研究基因组和基因组进化的方法，以及复杂性状的遗传分析。虽然淀粉样蛋白的形成和遗传过程由于其相关的病理而被历史地研究，但淀粉样蛋白型聚集体的正常生理作用正变得越来越受到重视，从酵母到哺乳动物的许多有益的淀粉样蛋白功能的例子。有趣的是，在聚集体清除中起作用的强效hsp100型解聚酶在动物基因组中不存在。因此，该基金的中心假设是，像Hsp100一类的广谱分解气体与动物多细胞进化不相容，因为蛋白质聚集和维持特定蛋白质的淀粉样蛋白形式是动物发育所必需的。与此一致的是，在秀丽隐杆线虫胚胎的早期发育过程中显示出惊人的淀粉样蛋白型聚集体的丰度。这项资助的目的是表征淀粉样蛋白聚集体在发育过程中的作用，并通过鉴定和表征发育表达的候选动物分解物来研究它们的调控。为了实现这一目标，研究计划利用秀丽隐杆线虫和酿酒酵母的互补和协同实验优势。实验将1)通过表征候选内源性动物解聚酶活性来分析发育中重要的淀粉样蛋白的功能和调控；2)表征秀丽隐杆线虫发育中关键的易聚集蛋白及其调控因子；3)确定秀丽隐杆线虫淀粉化蛋白聚集体的功能。总而言之，这笔拨款将研究淀粉样蛋白及其专用分解物如何在早期动物发育的关键事件中发挥新的调节作用，包括母体和合子合成蛋白的储存、繁殖和分布。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。

Non-Technical Abstract Enzymatic catalysts capable of disentangling potentially disease-causing protein aggregates have been proposed as a therapeutic solution to neurodegenerative disease. Given that all other forms of non-animal life already have these enzymes, it raises the question of why have humans lost the ability to express these potentially beneficial remedies? The answer to animal-specific loss of protein disentangling enzymes may lie in a potential beneficial role of protein aggregates that is incompatible with the presence of potent enzymes capable of destroying them. In this grant, the investigators will test this hypothesis in a comprehensive way by using the complementary features of two powerful genetic model organisms. Newly identified and potentially more subtle enzymatic regulators of protein aggregation will be identified using the yeast model system and then tested for regulation of the protein aggregates recently found in young, healthy embryos of the nematode worms. This grant will carry out a comprehensive examination of protein aggregates and their dedicated enzymatic regulators and seek to understand how they may serve as novel mechanisms to control key events in early animal development including the storage of key developmental proteins that can be released as required. To further connections between science and society, funds will also be used to develop new initiatives that expose middle school students, teachers and undergraduates to methods for studying genomes and genome evolution as well as the genetic analysis of complex traits.Technical AbstractAlthough processes of amyloid formation and inheritance are historically studied because of their associated pathologies, normal physiological roles of amyloid-type aggregates are becoming more appreciated with many examples of beneficial amyloid functions in species ranging from yeast to mammals. Interestingly, the potent Hsp100-type disaggregase, which plays a role in aggregate clearance, is absent in animal genomes. Thus, the central hypothesis of this grant is that broad-spectrum disaggregases like the Hsp100 class are incompatible with the evolution of animal multicellularity because protein aggregation and the maintenance of the amyloid form of specific proteins are required for animal development. Consistent with this, a surprising abundance of amyloid-type aggregates is shown during early development of C. elegans embryos. The objective of this grant is to characterize the role of amyloid aggregates during development and to investigate their regulation by identifying and characterizing developmentally expressed candidate animal disaggregases. To accomplish this objective, research plan capitalizes on complementary and synergistic experimental advantages of C. elegans and S. cerevisiae. Experiments will 1) analyze the function and regulation of developmentally important amyloids by characterizing candidate endogenous animal disaggregase activities, 2) characterize key aggregation-prone proteins and their regulators in C. elegans development and 3) determine the function of C. elegans amyloidized protein aggregates. Taken together, this grant will examine how amyloids and their dedicated disaggregases serve as novel regulators of key events in early animal development including storage, propagation and distribution of maternal and zygotically synthesized proteins.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.