Department of Microbiology, University of Pennsylvania Perelman School of Medicine, Philadelphia, Pennsylvania, United States of America.
PLoS Pathog. 2012;8(11):e1003021. doi: 10.1371/journal.ppat.1003021. Epub 2012 Nov 8.
Merkel cell polyomavirus (MCV or MCPyV) is the first human polyomavirus to be definitively linked to cancer. The mechanisms of MCV-induced oncogenesis and much of MCV biology are largely unexplored. In this study, we demonstrate that bromodomain protein 4 (Brd4) interacts with MCV large T antigen (LT) and plays a critical role in viral DNA replication. Brd4 knockdown inhibits MCV replication, which can be rescued by recombinant Brd4. Brd4 colocalizes with the MCV LT/replication origin complex in the nucleus and recruits replication factor C (RFC) to the viral replication sites. A dominant negative inhibitor of the Brd4-MCV LT interaction can dissociate Brd4 and RFC from the viral replication complex and abrogate MCV replication. Furthermore, obstructing the physiologic interaction between Brd4 and host chromatin with the chemical compound JQ1(+) leads to enhanced MCV DNA replication, demonstrating that the role of Brd4 in MCV replication is distinct from its role in chromatin-associated transcriptional regulation. Our findings demonstrate mechanistic details of the MCV replication machinery; providing novel insight to elucidate the life cycle of this newly discovered oncogenic DNA virus.
默克尔细胞多瘤病毒(MCV 或 MCPyV)是第一个明确与癌症相关的人类多瘤病毒。MCV 诱导致癌的机制和大部分 MCV 生物学仍在很大程度上未被探索。在这项研究中,我们证明了溴结构域蛋白 4(Brd4)与 MCV 大 T 抗原(LT)相互作用,并在病毒 DNA 复制中发挥关键作用。Brd4 的敲低抑制了 MCV 的复制,而重组 Brd4 可以挽救这种抑制。Brd4 在核内与 MCV LT/复制起始复合物共定位,并将复制因子 C(RFC)募集到病毒复制位点。Brd4-MCV LT 相互作用的显性负抑制剂可以使 Brd4 和 RFC 从病毒复制复合物中解离,并阻止 MCV 的复制。此外,用化学化合物 JQ1(+) 阻断 Brd4 与宿主染色质之间的生理相互作用会导致 MCV DNA 复制增强,表明 Brd4 在 MCV 复制中的作用与其在染色质相关转录调控中的作用不同。我们的研究结果阐明了 MCV 复制机制的详细机制;为阐明这种新发现的致癌 DNA 病毒的生命周期提供了新的见解。