A short SUMOylation tag modulates transcription factor activity.

IF 4 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY

Journal of Biological Chemistry Pub Date : 2025-10-08 DOI:10.1016/j.jbc.2025.110807

Antoine Y Bouchard,Anaïs J I Vivet,Valérie C Cabana,Chongyang Li,Pierre Thibault,Marc P Lussier,Sylvie Mader,Laurent Cappadocia

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引用次数: 0

Abstract

SUMOylation is a post-translational modification (PTM) that regulates multiple aspects of protein biology, including the activity of transcription factors such as p53. While strategies exist to decrease protein SUMOylation in a targeted manner, options are limited to increase SUMOylation in a protein-specific manner. Here, we developed a strategy to induce SUMOylation of a target protein relying on its genetic fusion to a 32-residue tag composed of the SUMO E3 module of ZNF451. Through in vitro and cell-based assays, we establish that this SUMOylation tag promotes robust poly/multi-SUMOylation of p53, used as a model substrate, with a strong preference for SUMO2/3 as compared to SUMO1. Mass spectrometry experiments performed on transfected HEK293 cells stably expressing a modified form of SUMO3 indicate that lysine 386, the main SUMOylation acceptor site of p53, is the primary target of ZNF-mediated SUMOylation. Increased SUMOylation represses p53 transcriptional activity in luciferase reporter assays, a result compatible with the general repressive effects of SUMOylation on transcription factor activity. Finally, fusion of ZNF to HSF1 and DNMT3A also increase their SUMOylation level, showcasing that ZNF could potentially be used to promote the SUMOylation of a broad range of proteins implicated in DNA metabolism. Overall, this strategy will facilitate the investigation of the impact of increased SUMOylation on specific protein substrates.

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短SUMOylation标签调节转录因子活性。

SUMOylation是一种翻译后修饰（PTM），可调节蛋白质生物学的多个方面，包括转录因子（如p53）的活性。虽然存在以靶向方式降低蛋白质summoylation的策略，但以蛋白质特异性方式增加summoylation的选择有限。在这里，我们开发了一种策略来诱导目标蛋白的summoylation依赖于其与由ZNF451的SUMO E3模块组成的32个残基标签的遗传融合。通过体外和基于细胞的实验，我们发现这个summoylation标签促进了p53的多/多summoylation，作为一个模型底物，与summo1相比，summo2 /3具有强烈的偏好。在稳定表达SUMO3修饰形式的转染HEK293细胞上进行的质谱实验表明，赖氨酸386是p53的主要sumo酰化受体位点，是znf介导的sumo酰化的主要靶点。在荧光素酶报告基因检测中，SUMOylation的增加抑制了p53的转录活性，这一结果与SUMOylation对转录因子活性的一般抑制作用相一致。最后，ZNF与HSF1和DNMT3A的融合也增加了它们的sumo化水平，表明ZNF可能被用来促进与DNA代谢有关的一系列蛋白质的sumo化。总的来说，这一策略将有助于研究增加的sumo酰化对特定蛋白质底物的影响。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Biological Chemistry Biochemistry, Genetics and Molecular Biology-Biochemistry

自引率

4.20%

发文量

1233

期刊介绍： The Journal of Biological Chemistry welcomes high-quality science that seeks to elucidate the molecular and cellular basis of biological processes. Papers published in JBC can therefore fall under the umbrellas of not only biological chemistry, chemical biology, or biochemistry, but also allied disciplines such as biophysics, systems biology, RNA biology, immunology, microbiology, neurobiology, epigenetics, computational biology, ’omics, and many more. The outcome of our focus on papers that contribute novel and important mechanistic insights, rather than on a particular topic area, is that JBC is truly a melting pot for scientists across disciplines. In addition, JBC welcomes papers that describe methods that will help scientists push their biochemical inquiries forward and resources that will be of use to the research community.