Department of Molecular Genetics and Microbiology, University of Florida, Gainesville, FL 32610, USA.
Virology. 2010 Apr 10;399(2):201-11. doi: 10.1016/j.virol.2010.01.012. Epub 2010 Feb 8.
An IPTG-inducible mutant in the E6R gene of vaccinia virus was used to study the role of the E6 virion core protein in viral replication. In the absence of the inducer, the mutant exhibited a normal pattern DNA replication, concatemer resolution and late gene expression, but it showed an inhibition of virion structural protein processing it failed to produce infectious particles. Electron microscopic analysis showed that in the absence of IPTG viral morphogenesis was arrested before IV formation: crescents, aberrant or empty IV-like structures, and large aggregated virosomes were observed throughout the cytoplasm. The addition of IPTG to release a 12-h block showed that virus infectious particles could be formed in the absence of de novo DNA synthesis. Our observations show that in the absence of E6 the association of viroplasm with viral membrane crescents is impaired.
我们利用痘苗病毒 E6R 基因的 IPTG 诱导突变体来研究 E6 衣壳核心蛋白在病毒复制中的作用。在没有诱导剂的情况下,突变体表现出正常的 DNA 复制、串联体分辨率和晚期基因表达模式,但它显示出病毒结构蛋白加工的抑制作用,无法产生感染性颗粒。电子显微镜分析显示,在没有 IPTG 的情况下,病毒形态发生在 IV 形成之前被阻断:新月形、异常或空的 IV 样结构,以及大的聚集的类病毒体在整个细胞质中被观察到。添加 IPTG 以释放 12 小时的阻断作用表明,在没有新的 DNA 合成的情况下,可以形成病毒感染性颗粒。我们的观察结果表明,在没有 E6 的情况下,类病毒体与病毒膜新月体的结合受到损害。