NSF-BSF: Control of molecular, cellular, and organismal phenotypes by the transcription factor NFAT5

该项目研究鱼类如何科普它们生活的水中盐所施加的压力。理解这一影响生存、生长和繁殖的重要过程的关键是研究鱼类如何感知环境盐度，以及它们如何产生内部信号，协调分子网络的调节，以抵消压力。该项目研究所涉及的分子及其相互作用，使用新的分子，遗传和生物化学工具结合定制的计算模型。通过研究对盐胁迫的反应，特别是，这个项目有可能帮助理解，一般来说，在不同层次的生物组织对环境压力的反应。通过研究罗非鱼（全球第二大水产养殖物种）的这些过程，这项工作将对基本食品供应系统产生积极影响。 此外，罗非鱼可以作为水生脊椎动物物种的应激生物学的模型系统。该项目产生的工具和知识通过相关的公共知识库和交流平台与更广泛的科学界分享。它包括精心设计的跨学科培训的研究人员在所有的职业生涯阶段，重点是代表性不足的少数民族，在尖端的实验和网络modeling approaches.This project identifies regulatory networks that are causally linked to molecular，cellular，and organismal phenotypes critical for salinity tolerance in tilapia，Oreochromis mossambicus.研究假设NFAT 5是一种转录因子，是控制这些网络的中央协调器。NFAT 5和适应性表型（耐受性，生长，繁殖力）之间的假定因果关系在盐度胁迫（高渗）将进行测试。该项目基于先前的研究，阐明了分子机制，并为研究鱼类适应性表型的机械原因提供了工具。NFAT 5功能将通过研究依赖于NFAT 5的分子网络和细胞和生物表型在系统规模，整体水平上确定。重点是将转录组和蛋白质组网络的高渗、NFAT 5依赖性调节与生长、存活、增殖和繁殖联系起来。将确定转录组与蛋白质组调控耦合程度的基本规则，并在不同的细胞/组织类型和基因组背景中测试其普遍性。细胞系将用于高通量基因靶向和细胞表型分析，以确定产生用于生物体表型分析的NFAT 5+和NFAT 5-转基因鱼的最佳靶标。更广泛的影响包括促进预测和减轻盐度胁迫对高商业价值生物的后果。一般的适应性策略来科普压力将被揭示和广泛分享。工具（工程细胞系，质粒，蛋白质组学文库）和产生的想法将通过GitHub等通信平台和会议共享。跨学科培训和推广将提供给研究生，REU学生，博士后，萨克拉门托地区社区学院的学生，K-12学生和教师，和公众。这个美国/以色列项目得到美国国家科学基金会和以色列两国科学基金会的支持。该奖项反映了NSF的法定使命，并通过使用基金会的智力价值进行评估而被认为值得支持和更广泛的影响审查标准。

This project investigates how fish cope with the stress imposed by salt in the water in which they live. Key to understanding this important process -- which impacts survival, growth, and reproduction -- is to examine how the fish sense environmental salinity and how they generate, in response, internal signals that orchestrate the regulation of molecular networks that counteract the stress. The project examines the molecules involved, and their interactions, using novel molecular, genetic, and biochemical tools in combination with custom-built computational models. By studying responses to salt stress, specifically, this project has the potential to help understand, in general, responses to environmental stress at different levels of biological organization. By examining these processes in tilapia, the second-most important aquaculture species globally, this work will positively impact an essential food supply system. In addition, tilapia can serve as a model system for stress biology in aquatic vertebrate species. This project generates tools and knowledge shared with the broader scientific community via pertinent public repositories and communication platforms. It includes elaborate cross-disciplinary training of researchers at all career stages, with emphasis on underrepresented minorities, in cutting-edge experimental and network modeling approaches.This project identifies regulatory networks that are causally linked to molecular, cellular, and organismal phenotypes critical for salinity tolerance in tilapia, Oreochromis mossambicus. The research hypothesis is that NFAT5, a transcription factor, is a central orchestrator for controlling these networks. Putative causal links between NFAT5 and adaptive phenotypes (tolerance, growth, fecundity) during salinity stress (hyperosmolality) will be tested. The project is based on prior research elucidating molecular mechanisms and generating tools for studying mechanistic causes of adaptive phenotypes in fish. NFAT5 functions will be determined on a systems-scale, holistic level by investigating molecular networks and cellular and organismal phenotypes that depend on NFAT5. The emphasis is on linking the hyperosmotic, NFAT5-dependent regulation of transcriptome and proteome networks with growth, survival, proliferation, and reproduction. Rules underlying the extent of coupling of transcriptome versus proteome regulation will be determined and their universality tested in different cell-/tissue types and genomic backgrounds. Cell lines will be utilized for high-throughput gene targeting and cellular phenotyping to identify the best targets for generating NFAT5+ and NFAT5- transgenic fish for organismal phenotyping. Broader impacts include the facilitation of prediction and mitigation of consequences of salinity stress on organisms of high commercial value. General adaptive strategies used to cope with stress will be revealed and shared broadly. Tools (engineered cell lines, plasmids, proteomics libraries) and ideas generated will be shared via communication platforms such as GitHub and at conferences. Cross-disciplinary training and outreach will be provided to graduate students, REU students, a postdoc, Sacramento area community college students, K-12 students and teachers, and the public.This US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.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.