TRIM proteins and the SUMO pathway: a systematic and functional analysis

TRIM 蛋白和 SUMO 途径：系统和功能分析

• 批准号: RGPIN-2017-06315
• 负责人: Berthoux, Lionel
• 金额: $ 1.89万
• 依托单位: Université du Québec à Trois-Rivières
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748772
• 关键词: TRIM; proteins; SUMO; pathway; systematic

TRIM proteins are a large family of proteins that share a common basic structure but are very diverse in their functions and subcellular localization. They harbor a domain called the RING finger which has an “E3 ubiquitin ligase” activity, meaning that it guides the protein modification process known as ubiquitination. Ubiquitination consists in the linkage of a small protein, ubiquitin, to other proteins. It is believed that their ubiquitin ligase activity is central to the function of TRIM proteins. Some TRIM proteins were found to be themselves subjected to another protein modification process known as SUMOylation. SUMOylation also consists in the linkage of a small protein, SUMO, to another protein and occurs at consensus sites. SUMOylation often modulates the intracellular localization, stability and/or function of the modified proteins. Interestingly, ubiquitination and SUMOylation are inter-dependent processes. Work in our laboratory showed that one human TRIM in particular, TRIM5, harbors a consensus SUMOylation site that modulates the ubiquitin ligase activity of the adjacent RING domain. Computer searches for SUMOylation consensus sites suggested that the majority of human TRIM proteins harbor such motifs, implying that SUMOylation plays an important and general role in the function of proteins of this family. We propose to establish a research program with the long-term goal of deciphering the functional relationships between TRIM proteins and SUMOylation. We will clone, express and purify all ~80 human TRIM proteins, including some that have never been studied. Their capacity to bind SUMO and to be SUMOylated will be investigated. SUMOylation will be tested in cells and using in vitro assays, and SUMOylation sites will be identified. The role of SUMOylation and SUMO-relevant motifs in the behavior of TRIM proteins will be assessed by analyzing their impact on the subcellular localization, stability and E3 ubiquitin ligase activity. This research program will yield the first detailed description of the TRIM proteins SUMOylation landscape and will uncover the role of SUMOylation in TRIM functions. Our systematic approach will allow us to uncover regulatory pathways common to multiple TRIM proteins and to characterize poorly studied members of the TRIM family. In addition to significantly advancing scientific knowledge, this research program has potential applications in protein engineering.

TRIM蛋白是一个大家族的蛋白质，它们具有共同的基本结构，但在功能和亚细胞定位方面非常多样。它们拥有一个称为RING finger的结构域，该结构域具有“E3泛素连接酶”活性，这意味着它指导称为泛素化的蛋白质修饰过程。泛素化是一种小蛋白质，泛素，与其他蛋白质的连接。据信，它们的泛素连接酶活性对TRIM蛋白的功能至关重要。发现一些TRIM蛋白本身经历另一种称为SUMO化的蛋白质修饰过程。SUMO化还包括小蛋白SUMO与另一种蛋白的连接，并发生在共有位点。SUMO化通常调节经修饰的蛋白质的细胞内定位、稳定性和/或功能。有趣的是，泛素化和SUMO化是相互依赖的过程。我们实验室的工作表明，一个人类TRIM，特别是TRIM 5，拥有一个共识SUMO化位点，调节泛素连接酶活性的相邻环结构域。计算机搜索SUMO化一致位点表明，大多数人TRIM蛋白含有这样的基序，这意味着SUMO化在该家族蛋白质的功能中起着重要和普遍的作用。我们建议建立一个研究计划，长期目标是破译TRIM蛋白和SUMO化之间的功能关系。我们将克隆、表达和纯化所有~80种人类TRIM蛋白，包括一些从未被研究过的蛋白。将研究它们结合SUMO和被SUMO化的能力。将在细胞中使用体外试验检测SUMO化，并鉴定SUMO化位点。SUMO化和SUMO相关基序在TRIM蛋白行为中的作用将通过分析其对亚细胞定位、稳定性和E3泛素连接酶活性的影响来评估。这项研究计划将首次详细描述TRIM蛋白SUMO化景观，并将揭示SUMO化在TRIM功能中的作用。我们的系统方法将使我们能够发现多种TRIM蛋白共同的调控途径，并表征TRIM家族研究不足的成员。除了显着推进科学知识，该研究计划在蛋白质工程中具有潜在的应用。