Regulation of protein multi-functionality by 3 UTRs

3个UTR对蛋白质多功能性的调节

• 批准号: 10330234
• 负责人: Christine Mayr
• 金额: $ 70.8万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-14 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10330234
• 关键词: 3' Untranslated Regions; Binding Proteins; CRISPR/Cas technology; Cell Death; Cells; Code; Cytoplasmic Granules; Cytosol; Development; Elements; Engineering; Environment; Genes; Goals; Length; Mediating; Messenger RNA; Methods; Organelles; Phase; Post-Translational Protein Processing; Protein Biosynthesis; Protein Isoforms; Proteins; Regulation; Research; Role; Therapeutic; Transcript; Translating; Translations; Untranslated Regions; migration; protein aggregation; protein complex; protein function; protein protein interaction; recruit; scaffold

Regulation of protein multi-functionality by 3′UTRs SUMMARY Many protein functions are mediated by protein complexes whose formation is often regulated by abundance as higher levels increase the chance to encounter an interaction partner. mRNAs contain a coding region that is translated into protein, but they also contain a 3′ untranslated region (3′UTR). In addition to regulation by abundance, my lab discovered that protein function can be regulated by 3′UTRs as during protein synthesis 3′UTRs mediate protein-protein interactions. 3′UTR-dependent protein complex assembly is mediated by the local translation environment. Each mRNA generates its own translation environment that consists of the proteins bound by the mRNA together with the recruited proteins. As a result, mRNA isoforms with alternative 3′UTRs – that often differ substantially in length – provide drastically different translation environments, and thus encode different protein functions. Currently, thousands of 3′UTR-dependent functions are unknown because they cannot be inferred from canonical protein functions. We have developed a method to systematically identify protein functions mediated by long 3′UTR isoforms of multi-UTR genes using a CRISPR-based approach. We will identify 3′UTRs that mediate so far unknown protein functions involved in the evasion of cell death, in the regulation of migration, and differentiation. We currently know of two ways to achieve 3′UTR-dependent functions. As described above, an mRNA that contains a long 3′UTR can generate its own translation environment. Moreover, mRNAs can use elements in their 3′UTRs to localize to pre-existing translation environments that are formed by phase-separated cytosolic compartments. Within these large cytosolic membraneless organelles the environment is generated by many mRNAs together with their recruited proteins. We discovered such a compartment called TIS granule network. We determined hundreds of enriched mRNAs and observed that usually only half of transcripts with the same 3′UTR localize to TIS granules. This implies that proteins can have alternative functions depending on whether they are translated in the cytosol or in TIS granules. Our goal is to investigate how proteins change their function when translated within TIS granules. To study TIS granule-dependent protein functions, we have engineered cells that are unable to assemble TIS granules. For candidates whose mRNAs are strongly enriched in TIS granules, we are investigating if translation in TIS granules controls the addition of post- translational modifications, the establishment of specific protein complexes, or if it suppresses protein aggregation. If successful, our research will reveal a widespread role of mRNA in the compartmentalization and physical scaffolding during translation. It will show how elements in 3′UTRs contribute to the diversification of protein function. In the long-term, it will facilitate the development of mRNA therapeutics where inclusion of specific 3′UTR elements allows mRNAs to encode proteins with more robust or alternative functions.

通过3'UTRS调节蛋白质多功能 概括 许多蛋白质功能是由蛋白质复合物介导的，蛋白质复合物通常由抽象调节 随着较高水平的增加，遇到互动伙伴的机会。 mRNA包含一个编码区域 被翻译成蛋白质，但它们还包含一个3'未翻译的区域（3'UTR）。除了通过 抽象性，我的实验室发现蛋白质功能可以像蛋白质合成期间受到3'UTR的调节 3'UTRS介导蛋白质 - 蛋白质相互作用。 3'UTR依赖性蛋白质复合物组装由 本地翻译环境。每个mRNA都会产生自己的翻译环境，由 由mRNA与募集的蛋白质结合的蛋白质。结果，具有替代性的mRNA同工型 3'Utrs - 通常长度不同 - 提供截然不同的翻译环境，并且 因此编码不同的蛋白质功能。目前，数千个3'UTR依赖性功能未知 因为它们不能从规范蛋白功能中推断出来。我们已经开发了一种方法 系统地识别使用A的长3'UTR同工型介导的蛋白质功能使用A 基于CRISPR的方法。我们将确定3'Utrs介导迄今涉及的未知蛋白功能 细胞死亡的演变，迁移的调节和分化。 我们目前知道实现3'UTR依赖功能的两种方法。如上所述，一个mRNA 包含长3'UTR可以生成自己的翻译环境。此外，mRNA可以在 它们的3'Utrs本地化为由相位分离的胞质形成的预先存在的翻译环境 车厢。在这些大型的胞质膜细胞器中，环境是由许多人产生的 mRNA以及其招募的蛋白质。我们发现了这样一个称为TIS颗粒网络的隔间。 我们确定了数百个富集的mRNA，并观察到通常只有一半的转录本具有相同的转录本 3'utr本地化于颗粒。这意味着蛋白质可以取决于是否具有替代功能 它们在细胞质或Tis颗粒中翻译。我们的目标是研究蛋白质如何改变他们的 在TIS颗粒中翻译时功能。为了研究颗粒依赖性的蛋白质功能，我们有 无法组装颗粒的工程细胞。对于mRNA强烈的候选人 富含颗粒的颗粒，我们正在研究颗粒中的翻译是否控制在后加入。 翻译修饰，特定蛋白质复合物的建立或抑制蛋白 聚合。 如果成功，我们的研究将揭示mRNA在分隔和物理化中的宽度作用 翻译过程中的脚手架。它将显示3'UTR中的元素如何促进蛋白质的多样化 功能。从长远来看，它将促进mRNA疗法的发展，其中包含特定 3'UTR元素允许mRNA用更健壮或替代功能编码蛋白质。