Arginine methylation by PRMT2 controls the functions of the actin nucleator Cobl. Hou et al.

examples of new original data - the file names refer to the respective figure panels in Hou et al Arginine methylation by PRMT2 controls the functions of the actin nucleator Cobl Wenya Hou1, Sabine Nemitz1, Simone Schopper2, Michael Lund Nielsen2, Michael Manfred Kessels1*, Britta Qualmann1* 1 Institute of Biochemistry I, Jena University Hospital - Friedrich Schiller University Jena, 07743 Jena, Germany 2 Novo Nordisk Foundation Center for Protein Research, University of Copenhagen, DK-2200 Copenhagen N, Denmark Lead contact [e-mail] * Corresponding authors: M. M. Kessels & B. Qualmann Institut für Biochemie I, UKJ, Nonnenplan 2-4, D-07743 Jena, Germany E-mail: [e-mail] [e-mail] Summary The complex architecture of neuronal networks in the brain requires tight control of the actin cytoskeleton. The actin nucleator Cobl is critical for neuronal morphogenesis. Here we unveil that Cobl is controlled by arginine methylation. Coprecipitations, coimmunoprecipitations, cellular reconstitutions and in vitro reconstitutions demonstrated that Cobl associates with the protein arginine methyltransferase PRMT2 in an SH3 domain-dependent manner and that this promotes methylation of Cobl’s actin nucleating C terminal domain. Consistently, PRMT2 phenocopied Cobl functions in both gain- and loss-of-function studies, both PRMT2- and Cobl-promoted dendritogenesis relied on methylation and PRMT2 required functionality of its catalytic and its SH3 domain. Furthermore, Cobl-mediated dendritic arborization required PRMT2, complex formation with PRMT2 and PRMT2’s catalytic activity. Mechanistic studies unveiled that Cobl methylation is key for Cobl’s actin binding. Therefore, arginine methylation is a regulatory mechanism reaching beyond controlling nuclear processes. It also controls a major, cytosolic, cytoskeletal component shaping neuronal cells.

新原创数据示例——本数据集的文件名称对应Hou等人发表的题为《PRMT2介导的精氨酸甲基化调控肌动蛋白成核因子Cobl功能》的研究论文（原文标题：Arginine methylation by PRMT2 controls the functions of the actin nucleator Cobl）

作者：侯文娅¹, 萨宾·内米茨¹, 西蒙娜·朔珀², 迈克尔·伦德·尼尔森², 迈克尔·曼弗雷德·凯塞尔¹*, 布丽塔·夸尔曼¹*

单位：
¹ 德国耶拿弗里德里希·席勒大学耶拿大学医院第一生物化学研究所，耶拿 07743
² 丹麦哥本哈根大学诺和诺德基金会蛋白质研究中心，哥本哈根N区 DK-2200

首席联系人：[e-mail]

* 共同通讯作者：M. M. 凯塞尔与B. 夸尔曼
德国耶拿UKJ第一生物化学研究所，Nonnenplan 2-4，邮编D-07743
电子邮箱：[e-mail]、[e-mail]

研究摘要
大脑内神经元网络的复杂架构，需要对肌动蛋白细胞骨架（actin cytoskeleton）进行严格调控。肌动蛋白成核因子（actin nucleator）Cobl对于神经元形态发生（neuronal morphogenesis）至关重要。本研究揭示，Cobl的功能受精氨酸甲基化调控。
本研究通过共沉淀实验、免疫共沉淀（coimmunoprecipitation）实验、细胞重构实验（cellular reconstitution）与体外重构实验（in vitro reconstitution）证实：Cobl可通过SH3结构域（SH3 domain）依赖的方式与蛋白精氨酸甲基转移酶2（Protein Arginine Methyltransferase 2, PRMT2）结合，且该结合可促进Cobl肌动蛋白成核C端结构域的甲基化修饰。功能获得与功能缺失实验均证实，PRMT2可重现Cobl的功能；PRMT2与Cobl共同介导的树突发生（dendritogenesis）过程均依赖甲基化修饰，且PRMT2需其催化结构域与SH3结构域发挥功能。此外，Cobl介导的树突分支形成（dendritic arborization）过程需要PRMT2、二者的复合物形成以及PRMT2的催化活性。机制研究进一步揭示，Cobl的甲基化修饰是其结合肌动蛋白的关键。
由此可见，精氨酸甲基化是一类突破核内调控范畴的调控机制，该修饰还可调控塑造神经元细胞的核心胞质细胞骨架组分。