Dallaire Frédéric, Schreiner Sabrina, Blair G Eric, Dobner Thomas, Branton Philip E, Blanchette Paola
Department of Biochemistry, McGill University, Montreal, Québec, Canada.
Institute of Virology, Technische Universität München/Helmholtz Zentrum München, Munich, Germany.
mSphere. 2015 Nov 11;1(1). doi: 10.1128/mSphere.00014-15. eCollection 2016 Jan-Feb.
The human adenovirus E4orf6/E1B55K E3 ubiquitin ligase is well known to promote viral replication by degrading an increasing number of cellular proteins that inhibit the efficient production of viral progeny. We report here a new function of the adenovirus 5 (Ad5) viral ligase complex that, although at lower levels, mimics effects of E1A products on E2F transcription factors. When expressed in the absence of E1A, the E4orf6 protein in complex with E1B55K binds E2F, disrupts E2F/retinoblastoma protein (Rb) complexes, and induces hyperphosphorylation of Rb, leading to induction of viral and cellular DNA synthesis as well as stimulation of early and late viral gene expression and production of viral progeny of E1/E3-defective adenovirus vectors. These new and previously undescribed functions of the E4orf6/E1B55K E3 ubiquitin ligase could play an important role in promoting the replication of wild-type viruses. IMPORTANCE During the course of work on the adenovirus E3 ubiquitin ligase formed by the viral E4orf6 and E1B55K proteins, we found, very surprisingly, that expression of these species was sufficient to permit low levels of replication of an adenovirus vector lacking E1A, the central regulator of infection. E1A products uncouple E2F transcription factors from Rb repression complexes, thus stimulating viral gene expression and cell and viral DNA synthesis. We found that the E4orf6/E1B55K ligase mimics these functions. This finding is of significance because it represents an entirely new function for the ligase in regulating adenovirus replication.
人腺病毒E4orf6/E1B55K E3泛素连接酶通过降解越来越多抑制病毒子代高效产生的细胞蛋白来促进病毒复制,这是广为人知的。我们在此报告腺病毒5(Ad5)病毒连接酶复合物的一项新功能,即尽管水平较低,但它能模拟E1A产物对E2F转录因子的作用。当在没有E1A的情况下表达时,与E1B55K复合的E4orf6蛋白结合E2F,破坏E2F/视网膜母细胞瘤蛋白(Rb)复合物,并诱导Rb的过度磷酸化,导致病毒和细胞DNA合成的诱导,以及早期和晚期病毒基因表达的刺激和E1/E3缺陷型腺病毒载体病毒子代的产生。E4orf6/E1B55K E3泛素连接酶的这些新的和以前未描述的功能可能在促进野生型病毒的复制中发挥重要作用。重要性在对由病毒E4orf6和E1B55K蛋白形成的腺病毒E3泛素连接酶的研究过程中,我们非常惊讶地发现,这些蛋白的表达足以使缺乏感染中心调节因子E1A的腺病毒载体进行低水平复制。E1A产物使E2F转录因子与Rb抑制复合物解偶联,从而刺激病毒基因表达以及细胞和病毒DNA合成。我们发现E4orf6/E1B55K连接酶模拟了这些功能。这一发现具有重要意义,因为它代表了连接酶在调节腺病毒复制方面的全新功能。