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.

3'UTR 对蛋白质多功能性的调节 概括 许多蛋白质功能是由蛋白质复合物介导的，其形成通常受丰度调节 因为较高的水平会增加遇到包含编码区的相互作用伙伴的机会。 被翻译成蛋白质，但除了受调控外，还含有 3' 非翻译区 (3'UTR)。 丰度，我的实验室发现蛋白质功能可以通过 3'UTR 进行调节，就像蛋白质合成过程中一样 3'UTR 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 3'UTR 介导。 每个 mRNA 都会生成自己的翻译环境，其中包括 蛋白质与 mRNA 结合在一起，从而产生具有替代性的 mRNA 亚型。 3'UTR——长度通常相差很大——提供了截然不同的翻译环境，并且 从而编码不同的蛋白质功能，目前，数以千计的 3'UTR 依赖性功能是未知的。 因为它们不能从典型的蛋白质功能中推断出来，我们已经开发了一种方法来推断。 必须使用多 UTR 基因的长 3'UTR 亚型介导的蛋白质功能 基于 CRISPR 的方法将鉴定介导迄今为止未知的蛋白质功能的 3'UTR。 在迁移和分化的调节中逃避细胞死亡。 目前我们知道有两种方法可以实现 3'UTR 依赖性功能。 包含长的3'UTR可以生成自己的翻译环境，而且mRNA可以使用其中的元件。 它们的 3'UTR 定位到由相分离的胞质形成的预先存在的翻译环境 在这些大的胞质无膜细胞器内，环境是由许多细胞器产生的。 我们发现了这样一个称为 TIS 颗粒网络的隔室。 我们测定了数百个富集的 mRNA，并观察到通常只有一半的转录物具有相同的 3'UTR 定位于 TIS 颗粒，这意味着蛋白质可以具有替代功能，具体取决于是否存在。 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它们的性质。 在 TIS 颗粒内翻译时的功能 为了研究 TIS 颗粒依赖性蛋白质功能，我们有 无法组装 TIS 颗粒的工程细胞。 富含 TIS 颗粒，我们正在研究 TIS 颗粒中的翻译是否控制后处理的添加 翻译修饰、特定蛋白质复合物的建立，或者是否抑制蛋白质 聚合。 如果成功，我们的研究将揭示 mRNA 在区划和物理方面的广泛作用。 它将展示 3'UTR 中的元素如何促进蛋白质的多样化。 从长远来看，它将促进包含特定功能的 mRNA 疗法的发展。 3'UTR 元件允许 mRNA 编码具有更强大或替代功能的蛋白质。