Binné Ulrich K, Amon Wolfgang, Farrell Paul J
Ludwig Institute for Cancer Research. Virology and Cell Biology Section, Imperial College Faculty of Medicine, St. Mary's Campus, Norfolk Place, London W2 1PG, United Kingdom.
J Virol. 2002 Oct;76(20):10282-9. doi: 10.1128/jvi.76.20.10282-10289.2002.
A luciferase reporter system with stably transfected oriP plasmids in Akata Burkitt's lymphoma cells provides a quantitative assay for the BZLF1 Zp promoter in response to B-cell receptor (BCR) activation by cross-linking with anti-immunoglobulin. In this system, detailed kinetic studies of promoter activity are possible. Previously reported promoter elements upstream of -221 from the transcription start and the ZIIR sequence had little effect on the Zp promoter, but the ZI and ZIIIA elements were essential for early activation. The ZIIIB element mediates autoactivation. Mutation of the ZV repressor sequence greatly increased the induction of the promoter but did not make it constitutively active. Zp transcription in response to BCR cross-linking declined after a few hours; this decline was reduced and delayed by acyclovir or phosphonoacetic acid, indicating that viral DNA replication or a late viral gene can play a role in the switch off of the Zp promoter. Late expression of the LMP1 protein may account for this.
在Akata伯基特淋巴瘤细胞中,带有稳定转染oriP质粒的荧光素酶报告系统可用于定量检测BZLF1 Zp启动子对通过抗免疫球蛋白交联激活B细胞受体(BCR)的反应。在该系统中,可以对启动子活性进行详细的动力学研究。先前报道的转录起始点上游-221处的启动子元件和ZIIR序列对Zp启动子影响很小,但ZI和ZIIIA元件对早期激活至关重要。ZIIIB元件介导自激活。ZV阻遏序列的突变极大地增加了启动子的诱导,但并未使其组成型激活。BCR交联后几小时,Zp转录下降;阿昔洛韦或膦甲酸可减少并延迟这种下降,表明病毒DNA复制或晚期病毒基因可能在Zp启动子的关闭中起作用。LMP1蛋白的晚期表达可能对此负责。