Nonkwelo C, Skinner J, Bell A, Rickinson A, Sample J
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
J Virol. 1996 Jan;70(1):623-7. doi: 10.1128/JVI.70.1.623-627.1996.
In Epstein-Barr virus (EBV)-transformed B lymphoblastoid and many Burkitt lymphoma cell lines, the EBV EBNA-1 protein is one of six viral nuclear antigens expressed from a common transcription unit under the control of one of two promoters, Wp or Cp. In contrast, EBNA-1 is the only EBV nuclear antigen expressed in Burkitt and other EBV-positive tumors. We previously identified a promoter of EBNA-1 transcription, designated Fp, in early-passage Mutu Burkitt tumor cells, and this promoter is also active in long-term Mutu and Akata Burkitt cell lines which maintain the exclusive expression of EBNA-1 characteristic of the tumor. However, transcription initiation within Fp reporter gene plasmids in EBV-negative cells occurs at positions 100 to 200 bases downstream of the Fp start site in the BamHI-Q restriction fragment. Here we demonstrate that transcription initiation within newly established Burkitt lymphoma cell lines is consistent with the transcription initiation we observed in reporter plasmids. Furthermore, previous observations of transcription from Fp to generate EBNA-1 transcripts can be attributed to lytic-cycle gene expression. These data, in conjunction with our previous characterization of promoter regulatory elements, define a fourth EBNA-1 promoter, Qp, that is active in latently infected Burkitt tumor cells.
在爱泼斯坦-巴尔病毒(EBV)转化的B淋巴母细胞和许多伯基特淋巴瘤细胞系中,EBV EBNA-1蛋白是由一个共同转录单元表达的六种病毒核抗原之一,该转录单元受两个启动子之一Wp或Cp的控制。相比之下,EBNA-1是在伯基特淋巴瘤和其他EBV阳性肿瘤中表达的唯一EBV核抗原。我们之前在早期传代的Mutu伯基特肿瘤细胞中鉴定出一个EBNA-1转录启动子,命名为Fp,并且该启动子在长期培养的Mutu和Akata伯基特细胞系中也具有活性,这些细胞系维持着肿瘤特有的EBNA-1的排他性表达。然而,在EBV阴性细胞中,Fp报告基因质粒内的转录起始发生在BamHI-Q限制片段中Fp起始位点下游100至200个碱基处。在这里,我们证明新建立的伯基特淋巴瘤细胞系中的转录起始与我们在报告质粒中观察到的转录起始一致。此外,之前关于从Fp转录产生EBNA-1转录本的观察结果可归因于裂解周期基因表达。这些数据,结合我们之前对启动子调控元件的表征,确定了第四个EBNA-1启动子Qp,它在潜伏感染的伯基特肿瘤细胞中具有活性。