Characterization of the interactions of human papillomavirus type 16 E6 with p53 and E6-associated protein in insect and human cells
Characterization of the interactions of human papillomavirus type 16 E6 with p53 and E6-associated protein in insect and human cells
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DOI：
10.1099/0022-1317-79-3-489
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发表时间：
1998-03-01
影响因子：
3.8
通讯作者：
Maitland, NJ
中科院分区：
文献类型：
--
作者：
Daniels, PR;Sanders, CM;Maitland, NJ
人乳头瘤病毒（HPV） 16e6通过泛素依赖的蛋白水解途径诱导肿瘤抑制蛋白p53的降解，在体外，这一过程涉及E6、p53和细胞蛋白E6相关蛋白（E6- ap）之间形成三分子复合物，然而，他们在体内的潜在相互作用尚未进行分析，我们建立了一个模型来表达和分析这三种蛋白在昆虫细胞中的相互作用。一种真核系统，在那里蛋白质可能发生关键性的修饰。在杆状病毒感染的细胞中，p53的降解可以发生，然而，由于后期缺乏ATP， p53仅在感染周期的早期降解。因此，在这个系统中可以产生大量的材料进行进一步的分析。也有证据表明，在体内，E6可以在没有EG-AP的情况下与p53相互作用，而EG-AP可以在没有E6的情况下与p53相互作用。此外，利用生化分离和间接免疫荧光对蛋白质的亚细胞定位分析表明，p53的降解发生在细胞的核周区域。
Human papillomavirus (HPV) 16 E6 induces the degradation of the tumour suppressor protein p53 by the ubiquitin-dependent proteolysis pathway, In vitro, this process involves the formation of a trimolecular complex between E6, p53 and a cellular protein E6-associated protein (E6-AP), However, an analysis of their potential interactions in vivo has not been carried out, We have established a model for the expression and analysis of the interactions of these three proteins in insect cells, a eukaryotic system where potentially crucial modifications of the proteins will occur. In baculovirus-infected cells the degradation of p53 can occur, However, p53 is only degraded early in the infectious cycle due to a lack of ATP at later times. Consequently, substantial quantities of material can be produced in this system for further analysis, Evidence is also provided that, in vivo, E6 can interact with p53 in the absence of EG-AP and that EG-AP can interact with p53 in the absence of E6. Furthermore, analysis of the subcellular localization of the proteins using both biochemical fractionation and indirect immunofluorescence suggests that the degradation of p53 occurs in the perinuclear region of the cell.