de Boor Susanne, Knyphausen Philipp, Kuhlmann Nora, Wroblowski Sarah, Brenig Julian, Scislowski Lukas, Baldus Linda, Nolte Hendrik, Krüger Marcus, Lammers Michael
Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, 50931 Cologne, Germany.
Institute for Genetics and Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases, University of Cologne, 50931 Cologne, Germany
Proc Natl Acad Sci U S A. 2015 Jul 14;112(28):E3679-88. doi: 10.1073/pnas.1505995112. Epub 2015 Jun 29.
Ran is a small GTP-binding protein of the Ras superfamily regulating fundamental cellular processes: nucleo-cytoplasmic transport, nuclear envelope formation and mitotic spindle assembly. An intracellular Ran•GTP/Ran•GDP gradient created by the distinct subcellular localization of its regulators RCC1 and RanGAP mediates many of its cellular effects. Recent proteomic screens identified five Ran lysine acetylation sites in human and eleven sites in mouse/rat tissues. Some of these sites are located in functionally highly important regions such as switch I and switch II. Here, we show that lysine acetylation interferes with essential aspects of Ran function: nucleotide exchange and hydrolysis, subcellular Ran localization, GTP hydrolysis, and the interaction with import and export receptors. Deacetylation activity of certain sirtuins was detected for two Ran acetylation sites in vitro. Moreover, Ran was acetylated by CBP/p300 and Tip60 in vitro and on transferase overexpression in vivo. Overall, this study addresses many important challenges of the acetylome field, which will be discussed.
Ran是Ras超家族中的一种小GTP结合蛋白，可调节基本的细胞过程：核质运输、核膜形成和有丝分裂纺锤体组装。由其调节因子RCC1和RanGAP的不同亚细胞定位产生的细胞内Ran•GTP/Ran•GDP梯度介导了其许多细胞效应。最近的蛋白质组学筛选在人类中鉴定出5个Ran赖氨酸乙酰化位点，在小鼠/大鼠组织中鉴定出11个位点。其中一些位点位于功能上非常重要的区域，如开关I和开关II。在这里，我们表明赖氨酸乙酰化会干扰Ran功能的重要方面：核苷酸交换和水解、Ran的亚细胞定位、GTP水解以及与输入和输出受体的相互作用。在体外检测到某些去乙酰化酶对两个Ran乙酰化位点的去乙酰化活性。此外，Ran在体外以及体内转移酶过表达时被CBP/p300和Tip60乙酰化。总体而言，本研究解决了乙酰化蛋白质组领域的许多重要挑战，将对此进行讨论。
