Mechanistic Analysis of Papillimavirus E2 Functions

乳头瘤病毒(PV)是一组小的DMA病毒，可引起多种组织的良性病变。 脊椎动物，包括人类。某些乳头状瘤病毒，包括人乳头瘤病毒(HPV)，如 与HPV16和HPV18一样，也与宫颈癌和其他人类肿瘤的发病机制有关。 乳头瘤病毒建立持续的感染，在这种感染中，病毒基因组保持自主 在增殖的宿主细胞的细胞核中以低拷贝数复制质粒。在牛乳头瘤病毒中 (BPV1)转化的小鼠细胞，病毒基因组也以多拷贝核质粒的形式复制，可以 持续较长时间，保持细胞的转化状态。要确保持久化 无论是被感染的细胞还是转化的细胞，复制的病毒基因组都必须被分割并在 有丝分裂后的子细胞的核。没有机制来确保病毒的本地化 核膜内的基因组在核重组后，病毒基因组可能会留在 细胞分裂后由于降解或稀释而丢失的细胞质。乳头瘤病毒E2 蛋白质，这是一种重要的调节蛋白，在病毒转录和病毒DNA中发挥关键作用 复制，也是在分裂细胞中维持病毒基因组所必需的。E2，以及PV基因组， 在细胞分裂过程中与有丝分裂染色体密切相关。利用蛋白质组学方法 在体内系统地表征与E2相关的细胞蛋白，我们最近发现了Brd4 (含溴结构域的蛋白4)作为一种E2相互作用蛋白，并表明它具有有丝分裂的功能 BPV1E2的染色体受体。这项研究拨款的重点是延长这些关于光伏E2的研究 并研究了E2与Brd4相互作用对蛋白质其他方面的功能意义。 病毒的生命周期。破坏E2与Brd4的结合可以阻断BPV1和BPV1对细胞的转化 E_2和病毒DNA与有丝分裂染色体的关系将进行化学基因筛查 寻找E2/Brd4结合的小分子抑制剂作为新型治疗药物的潜在先导分子 抗病毒化合物。

The papillomaviruses (PVs) are a group of small DMAviruses that induce benign lesions in variety of higher vertebrates, including humans. Certain papillomaviruses, including the human papillomaviruses (HPVs) such as HPV16 and 18, are also associated with the pathogenesis of cervical cancer and other human tumors. Papillomaviruses establish persistent infections in which the viral genomes are maintained as autonomous replicating plasmids at a low copy number in the nuclei of proliferating host cells. In bovine papillomavirus (BPV1) transformed mouse cells, the viral genomes also replicate as multicopy nuclear plasmids that can persist over long periods of time to maintain the transformation status of the cells. To ensure persistence in both infected and transformed cells, the replicated viral genomes must be partitioned and maintained in the nuclei of daughter cells following mitosis. Without a mechanism to ensure the localization of the viral genome within the nuclear envelope following nuclear reassembly, the viral genome could be left behind in the cytoplasm and lost either through degradation or dilution after cell division. The papillomavirus E2 protein, which is an important regulatory protein that plays critical roles in viral transcriptional and viral DNA replication, is also required for viral genome maintenance in dividing cells. E2, as well as the PV genomes, are closely associated with mitotic chromosomes in dividing cells. Utilizing a proteomic approach to systematically characterize cellular proteins that associate with E2 in vivo, we recently identified Brd4 (bromodomain-containing protein 4) as an E2 interacting protein and showed that it functions as the mitotic chromosome receptor for BPV1 E2. The focus of this research grant is to extend these studies on the PV E2 proteins and to examine the functional significance of the interaction of E2 with Brd4 to other aspects of the viral life cycle. Disruption of the binding of E2 to Brd4 can block the transformation of cells by BPV1 and the association of E2 and the viral DNA with mitotic chromosomes. Chemical genetic screens will be conducted to identify small molecule inhibitors of E2/Brd4 binding as potential lead molecules for novel therapeutic antiviral compounds.