Plk1 interacts with RNF2 and promotes its ubiquitin-dependent degradation
Plk1 interacts with RNF2 and promotes its ubiquitin-dependent degradation
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DOI：
10.3892/or.2018.6326
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发表时间：
2018-05-01
期刊：
影响因子：
4.2
通讯作者：
Du, Jian
中科院分区：
文献类型：
--
作者：
An, Ran;Cheng, Li;Du, Jian
Ring finger protein 2（RNF2），也称为RING 2或RING 1B，在不同类型的癌症中显示致癌功能，然而，RNF2在有丝分裂期间的功能尚未被评估。酵母双杂交筛选进行了使用人类HeLa cDNA文库，以探索和鉴定与RNF2相互作用的蛋白质。几个阳性克隆，包括Polo样激酶1（Plk1），有丝分裂的关键调节，被确定。RNF2和Plk1之间的相互作用，证实使用α-半乳糖苷酶和生长试验中的选择性培养基，在体外谷胱甘肽S-转移酶下拉，和在体内免疫沉淀试验。此外，我们证实，RNF2共定位与Plk1在有丝分裂染色体的前中期和中期使用免疫荧光测定。此外，我们的研究结果表明，Plk1激酶活性所需的泛素依赖性降解RNF2。这些发现为理解RNF2在有丝分裂调控和肿瘤发生中的作用提供了新的线索。
Ring finger protein 2 (RNF2), also known as RING2 or RING1B, displays oncogenic functions in different types of cancers, yet, the function of RNF2 during mitosis has not been evaluated. A yeast two-hybrid screen was undertaken using a human HeLa cDNA library to explore and identify proteins that interact with RNF2. Several positive clones, including Polo-like kinase 1 (Plk1), a critical regulator of mitosis, were identified. The interaction between RNF2 and Plk1 was confirmed using a -galactosidase and growth test in selective media, in vitro glutathione S-transferase pull-down, and in vivo immunoprecipitation assays. Moreover, we confirmed that RNF2 co-localized with Plk1 at mitotic chromosomes in the prometaphase and metaphase using an immunofluorescence assay. In addition, our results revealed that Plk1 kinase activity was required for ubiquitin-dependent degradation of RNF2. These findings provide a new clue for understanding the function of RNF2 during mitotic regulation and tumorigenesis.