Zheng Qiaoli, Liu Hao, Ye Jingjia, Zhang Hui, Jia Zhenyu, Cao Jiang
Clinical Research Center, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China.
Institute of Occupational Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310007, P.R. China.
Mol Med Rep. 2016 Apr;13(4):2973-80. doi: 10.3892/mmr.2016.4935. Epub 2016 Feb 23.
Eukaryotic translation initiation factor 3 subunit g (eIF3g) is a core subunit of the eukaryotic translation initiation factor 3 complex, and is important in the initiation of translation. It is also involved in caspase-mediated apoptosis, and is upregulated in multidrug-resistant cancer cells. In the present study, the nuclear distribution of eIF3g was determined by performing co-immunoprecipitation of proteins that potentially interact with eIF3g in the nucleus. Mass spectrometry characterization showed that three proteins, heterogeneous nuclear ribonucleoprotein U/scaffold attachment factor A, HSZFP36/zinc finger protein 823 and β‑actin, were among the candidate eIF3g‑interacting proteins in the nucleus. The protein‑protein interaction was further confirmed by cross‑linking and a glutathione S‑transferase pull‑down assay, followed by western blotting. The co‑localization of these proteins was determined by confocal microscopy. These findings provide novel insight into the possible functions of eIF3g in the nucleus and serves as an important first step for further investigation of the roles of eIF3g in cancer development.
真核生物翻译起始因子3亚基g(eIF3g)是真核生物翻译起始因子3复合物的核心亚基,在翻译起始过程中起重要作用。它还参与半胱天冬酶介导的细胞凋亡,并且在多药耐药癌细胞中上调。在本研究中,通过对细胞核中可能与eIF3g相互作用的蛋白质进行共免疫沉淀来确定eIF3g的核分布。质谱表征显示,三种蛋白质,即不均一核核糖核蛋白U/支架附着因子A、HSZFP36/锌指蛋白823和β-肌动蛋白,是细胞核中候选的与eIF3g相互作用的蛋白质。通过交联和谷胱甘肽S-转移酶下拉试验进一步证实了蛋白质-蛋白质相互作用,随后进行蛋白质印迹分析。通过共聚焦显微镜确定这些蛋白质的共定位。这些发现为eIF3g在细胞核中的可能功能提供了新的见解,并作为进一步研究eIF3g在癌症发展中作用的重要第一步。