RoL:NSF-BSF: IMAGiNE: Temperature acclimation through RNA editing

RoL:NSF-BSF: IMAGiNE: 通过 RNA 编辑实现温度适应

• 批准号: 2110074
• 负责人: Joshua Rosenthal
• 金额: $ 114.01万
• 依托单位: Marine Biological Laboratory
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2110074&HistoricalAwards=false
• 关键词: RoL; NSF; BSF; IMAGiNE; Temperature

All the information required to create an organism, and for the organism to sustain its life, is encoded in its DNA. The portion of the information required at any given moment is temporarily copied into RNA before being decoded into the proteins that execute required functions. Molecular systems that manipulate this flow of information have led to some of the most consequential tools for modern biology, from gene cloning technologies to CRISPR genome engineering. All multicellular animals can edit genetic information as it passes through RNA, however the extent to which they do so varies widely. The coleoid cephalopods (squid, octopus, and cuttlefish) edit RNA at exceptionally high levels, and thus provide ideal models to study the process. Because RNA is a transient molecule, RNA editing is a good tool to create temporary changes that help with environmental acclimation. The work in this grant seeks to better understand how cephalopod RNA editing is used for temperature acclimation. Using recently developed cephalopod gene knockout technologies, the enzyme(s) that generate high-level RNA editing will be uncovered. In addition, how individual RNA edits affect proteins will be studied. Finally, the evolutionary underpinnings of RNA editing will be compared between similar squid species that inhabit different thermal environments. This project will enhance STEM education for underrepresented minorities through hands-on workshops on genome editing given to students from the University of Puerto Rico. Outreach on cephalopod biology and RNA editing will be extended to science journalists, and rabbinic leaders from Jewish community.Most organisms are confronted by a variable thermal environment and temperatures fluctuations are particularly acute for aquatic ectotherms due to the high thermal conductivity of water. All enzymatic reactions are governed by temperature, but the degree of sensitivity varies. Coordinating multiple enzymatic reactions across temperatures is a fundamental physiological challenge. This proposal focuses on RNA editing through adenosine deamination and how it is used for temperature acclimation. All multicellular metazoans use this process, however the coleoid cephalopods use it most extensively. Transcriptome-wide screens have uncovered tens of thousands of RNA editing sites, and these large data sets provide a unique opportunity to examine how editing is used to fine-tune physiology, and how it evolves. Taking advantage of recently developed gene knockout methods for cephalopods, the enzymatic underpinnings of high-level recoding will be explored, as well as the molecular basis of its temperature sensitivity. In addition, the work will investigate how temperature sensitive editing sites affect the function of key proteins in neurophysiology and examine how these sites are selected in closely related species from different thermal environments. This project will enhance STEM education for underrepresented minorities, generate resources for cephalopod biology, and provide diverse community outreach. Workshops on genome editing will be given to students from the University of Puerto Rico. The PIs will create guidelines for the humane and ethical use of cephalopods in research. Outreach on cephalopod biology and RNA editing will be extended to science journalists, and rabbinic leaders from Jewish community.This proposal was supported by the Integrative Ecological Physiology Program within the Division of Integrative Organismal Systems, the Genetic Mechanisms Cluster within the Division of Molecular and Cellular Biosciences, and Rules of Life Program within the Directorate for Biological Sciences.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.

创造一个有机体所需的所有信息，以及有机体维持其生命所需的所有信息，都编码在其DNA中。在任何给定时刻所需的信息部分在被解码成执行所需功能的蛋白质之前暂时复制到RNA中。操纵这种信息流的分子系统已经为现代生物学带来了一些最重要的工具，从基因克隆技术到CRISPR基因组工程。所有多细胞动物都可以在遗传信息通过RNA时对其进行编辑，但它们这样做的程度差异很大。鞘翅目头足类动物（鱿鱼、章鱼和乌贼）的RNA编辑水平非常高，因此为研究这一过程提供了理想的模型。由于RNA是一种瞬时分子，RNA编辑是一种很好的工具，可以创造有助于环境适应的临时变化。这项研究旨在更好地了解头足类动物RNA编辑如何用于温度适应。利用最近开发的头足类基因敲除技术，将发现产生高水平RNA编辑的酶。此外，还将研究单个RNA编辑如何影响蛋白质。最后，RNA编辑的进化基础将在居住在不同热环境的类似鱿鱼物种之间进行比较。该项目将通过为波多黎各大学的学生举办基因组编辑实践讲习班，加强对代表性不足的少数群体的STEM教育。头足类生物学和RNA编辑的推广将扩展到科学记者和犹太社区的拉比领导人。大多数生物都面临着多变的热环境，由于水的高导热性，水生外温动物的温度波动特别剧烈。所有酶促反应都受温度控制，但敏感程度不同。在不同温度下协调多个酶促反应是一项基本的生理挑战。该提案的重点是通过腺苷脱氨基进行RNA编辑以及如何将其用于温度适应。所有的多细胞后生动物都使用这个过程，但是最广泛使用的是鞘翅目头足类。转录组范围的屏幕已经发现了数万个RNA编辑位点，这些大型数据集提供了一个独特的机会来研究编辑如何用于微调生理学，以及它如何进化。利用最近开发的头足类基因敲除方法，将探讨高水平重新编码的酶基础，以及其温度敏感性的分子基础。此外，这项工作还将研究温度敏感的编辑位点如何影响神经生理学中关键蛋白质的功能，并研究这些位点如何在来自不同热环境的密切相关物种中被选择。该项目将加强代表性不足的少数民族的STEM教育，为头足类生物学提供资源，并提供多样化的社区外展。将为波多黎各大学的学生举办基因组编辑讲习班。PI将为头足类动物在研究中的人道和道德使用制定指导方针。头足类生物学和RNA编辑的推广将扩展到科学记者和犹太社区的拉比领导人。这一提议得到了综合有机系统部内的综合生态生理学计划，分子和细胞生物科学部内的遗传机制集群，该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识产权进行评估来支持。优点和更广泛的影响审查标准。

项目成果

Temperature-dependent RNA editing in octopus extensively recodes the neural proteome

• DOI: 10.1016/j.cell.2023.05.004
• 发表时间: 2023-06-08
• 期刊: CELL
• 影响因子: 64.5
• 作者: Birk, Matthew A.;Liscovitch-Brauer, Noa;Rosenthal, Joshua J. C.
• 通讯作者: Rosenthal, Joshua J. C.

Extensive Recoding of the Neural Proteome in Cephalopods by RNA Editing

• DOI: 10.1146/annurev-animal-060322-114534
• 发表时间: 2023-01-01
• 期刊: ANNUAL REVIEW OF ANIMAL BIOSCIENCES
• 影响因子: 12
• 作者: Rosenthal，Joshua J. C.;Eisenberg，Eli
• 通讯作者: Eisenberg，Eli