The Department of Medicine, Division of Infectious Disease, Brigham and Women's Hospital, Boston, Massachusetts, United States of America.
PLoS One. 2011;6(8):e22929. doi: 10.1371/journal.pone.0022929. Epub 2011 Aug 11.
Epstein Barr Virus (EBV) replicates in oral epithelial cells and gains entry to B-lymphocytes. In B-lymphocytes, EBV expresses a restricted subset of genes, the Latency III program, which converts B-lymphocytes to proliferating lymphoblasts. Latent Membrane Protein 1 (LMP1) and the other Latency III associated proteins are also expressed during virus replication. LMP1 is essential for virus replication and egress from Akata Burkitt Lymphoma cells, but a role in epithelial cell replication has not been established. Therefore, we have investigated whether LMP1 enhances EBV replication and egress from HEK293 cells, a model epithelial cell line used for EBV recombinant molecular genetics. We compared wild type (wt) and LMP1-deleted (LMP1Δ) EBV bacterial artificial chromosome (BAC) based virus replication and egress from HEK293. Following EBV immediate early Zta protein induction of EBV replication in HEK293 cells, similar levels of EBV proteins were expressed in wt- and LMP1Δ-infected HEK293 cells. LMP1 deletion did not impair EBV replication associated DNA replication, DNA encapsidation, or mature virus release. Indeed, virus from LMP1Δ-infected HEK293 cells was as infectious as EBV from wt EBV infected HEK cells. Trans-complementation with LMP1 reduced Rta expression and subsequent virus production. These data indicate that LMP1 is not required for EBV replication and egress from HEK293 cells.
爱泼斯坦-巴尔病毒(EBV)在口腔上皮细胞中复制,并进入 B 淋巴细胞。在 B 淋巴细胞中,EBV 表达一组受限的基因,即潜伏 III 程序,该程序将 B 淋巴细胞转化为增殖的淋巴母细胞。潜伏膜蛋白 1(LMP1)和其他潜伏 III 相关蛋白也在病毒复制过程中表达。LMP1 对病毒复制和从 Akata 伯基特淋巴瘤细胞中逸出是必需的,但在上皮细胞复制中的作用尚未确定。因此,我们研究了 LMP1 是否增强 EBV 在 HEK293 细胞(用于 EBV 重组分子遗传学的上皮细胞系模型)中的复制和逸出。我们比较了野生型(wt)和 LMP1 缺失(LMP1Δ)EBV 细菌人工染色体(BAC)基于的病毒复制和从 HEK293 细胞中的逸出。在 Zta 蛋白诱导 EBV 在 HEK293 细胞中复制后,wt-和 LMP1Δ 感染的 HEK293 细胞中表达了相似水平的 EBV 蛋白。LMP1 缺失不损害 EBV 复制相关的 DNA 复制、DNA 包裹或成熟病毒释放。事实上,来自 LMP1Δ 感染的 HEK293 细胞的病毒与来自 wt EBV 感染的 HEK 细胞的病毒一样具有感染力。LMP1 的反式互补减少了 Rta 的表达和随后的病毒产生。这些数据表明,LMP1 不是 EBV 从 HEK293 细胞中复制和逸出所必需的。