Lytic cycle gene regulation of Epstein-Barr virus
Lytic cycle gene regulation of Epstein-Barr virus
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DOI：
10.1128/jvi.78.24.13460-13469.2004
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发表时间：
2004-12-01
影响因子：
5.4
通讯作者：
Farrell, PJ
中科院分区：
文献类型：
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作者：
Amon, W;Binné, UK;Farrell, PJ
使用含有稳定转染到Akata Burkitt淋巴瘤细胞中的Epstein-Barr病毒（EBV）oriP序列的附加型报告质粒来分析EBV裂解周期基因调控。首先，我们发现EBV的Zp启动子，而不是Rp启动子，可以在这些oriP质粒中蛋白质合成的情况下被激活，从而对Rp的即时早期状态产生怀疑。Zp的额外水平的调节暗示了ZV元件的突变的分析。第二，我们对晚期裂解周期启动子的分析表明，正确的相对时间，依赖于顺式的orilyt，以及对DNA复制抑制剂的敏感性在oriP质粒上重建。晚期启动子荧光素酶活性的oriP质粒也纳入复制能力orilyt是膦酰乙酸敏感，EBV晚期基因的标志。一个最小的ori lyt，它只复制弱，是足以赋予晚的表达时间特异性晚启动子。最后，转录起始位点上游的EBV晚期启动子序列的缺失分析证实，-49和+30之间的序列对于晚期基因表达是足够的，这取决于顺式的ori lyt。然而，在许多晚期基因中发现的TATA盒的TATT版本对于晚期表达不是必需的。
Episomal reporter plasmids containing the Epstein-Barr virus (EBV) oriP sequence stably transfected into Akata Burkitt's lymphoma cells were used to analyze EBV lytic cycle gene regulation. First, we found that the Zp promoter of EBV, but not the Rp promoter, can be activated in the absence of protein synthesis in these oriP plasmids, casting doubt on the immediate early status of Rp. An additional level of regulation of Zp was implied by analysis of a mutation of the ZV element. Second, our analysis of late lytic cycle promoters revealed that the correct relative timing, dependence on ori lyt in cis, and sensitivity to inhibitors of DNA replication were reconstituted on the oriP plasmids. Late promoter luciferase activity from oriP plasmids also incorporating replication-competent ori lyt was phosphonoacetic acid sensitive, a hallmark of EBV late genes. A minimal ori lyt, which only replicates weakly, was sufficient to confer late timing of expression specifically on late promoters. Finally, deletion analysis of EBV late promoter sequences upstream of the transcription start site confirmed that sequences between -49 and +30 are sufficient for late gene expression, which is dependent on ori lyt in cis. However, the TATT version of the TATA box found in many late genes was not essential for late expression.