Zhang Dongyun, Liang Yuguang, Xie Qipeng, Gao Guangxun, Wei Jinlong, Huang Haishan, Li Jingxia, Gao Jimin, Huang Chuanshu
Nelson Institute of Environmental Medicine, New York University School of Medicine, Tuxedo, New York 10987 and; Zhejiang Provincial Key Laboratory for Technology and Application of Model Organisms, School of Life Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China.
Nelson Institute of Environmental Medicine, New York University School of Medicine, Tuxedo, New York 10987 and.
J Biol Chem. 2015 Feb 20;290(8):4784-4800. doi: 10.1074/jbc.M114.598219. Epub 2015 Jan 5.
Nucleolin is a ubiquitously expressed protein and participates in many important biological processes, such as cell cycle regulation and ribosomal biogenesis. The activity of nucleolin is regulated by intracellular localization and post-translational modifications, including phosphorylation, methylation, and ADP-ribosylation. Small ubiquitin-like modifier (SUMO) is a category of recently verified forms of post-translational modifications and exerts various effects on the target proteins. In the studies reported here, we discovered SUMOylational modification of human nucleolin protein at Lys-294, which facilitated the mRNA binding property of nucleolin by maintaining its nuclear localization. In response to arsenic exposure, nucleolin-SUMO was induced and promoted its binding with gadd45α mRNA, which increased gadd45α mRNA stability and protein expression, subsequently causing GADD45α-mediated cell death. On the other hand, ectopic expression of Mn-SOD attenuated the arsenite-generated superoxide radical level, abrogated nucleolin-SUMO, and in turn inhibited arsenite-induced apoptosis by reducing GADD45α expression. Collectively, our results for the first time demonstrate that nucleolin-SUMO at K294R plays a critical role in its nucleus sequestration and gadd45α mRNA binding activity. This novel biological function of nucleolin is distinct from its conventional role as a proto-oncogene. Therefore, our findings here not only reveal a new modification of nucleolin protein and its novel functional paradigm in mRNA metabolism but also expand our understanding of the dichotomous roles of nucleolin in terms of cancer development, which are dependent on multiple intracellular conditions and consequently the appropriate regulations of its modifications, including SUMOylation.
核仁素是一种广泛表达的蛋白质,参与许多重要的生物学过程,如细胞周期调控和核糖体生物合成。核仁素的活性受细胞内定位和翻译后修饰的调节,包括磷酸化、甲基化和ADP核糖基化。小泛素样修饰物(SUMO)是最近证实的一类翻译后修饰形式,对靶蛋白具有多种作用。在本研究中,我们发现人核核核核仁素蛋白在赖氨酸294位点发生SUMO化修饰,该修饰通过维持其核定位促进了核仁素与mRNA的结合特性。在暴露于砷的情况下,核仁素-SUMO被诱导并促进其与gadd45α mRNA的结合,从而增加gadd45α mRNA的稳定性和蛋白质表达,随后导致GADD45α介导的细胞死亡。另一方面,锰超氧化物歧化酶(Mn-SOD)的异位表达降低了亚砷酸盐产生的超氧自由基水平,消除了核仁素-SUMO,进而通过降低GADD45α表达抑制了亚砷酸盐诱导的细胞凋亡。总的来说,我们的结果首次表明,K294R位点的核仁素-SUMO在其核隔离和gadd45α mRNA结合活性中起关键作用。核仁素的这种新生物学功能不同于其作为原癌基因的传统作用。因此,我们的研究结果不仅揭示了核仁素蛋白的一种新修饰及其在mRNA代谢中的新功能模式,而且扩展了我们对核仁素在癌症发展中二分作用的理解,这取决于多种细胞内条件以及对其修饰(包括SUMO化)的适当调控。