Hohn Oliver, Hanke Kirsten, Lausch Veronika, Zimmermann Anja, Mostafa Saeed, Bannert Norbert
HIV and Other Retroviruses, Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany.
Viruses. 2014 Nov 11;6(11):4332-45. doi: 10.3390/v6114332.
The HERV-K(HML-2) family contains the most recently integrated and best preserved endogenized proviral sequences in the human genome. All known elements have nevertheless been subjected to mutations or deletions that render expressed particles non-infectious. Moreover, these post-insertional mutations hamper the analysis of the general biological properties of this ancient virus family. The expression of consensus sequences and sequences of elements with reverted post-insertional mutations has therefore been very instrumental in overcoming this limitation. We investigated the particle morphology of a recently reconstituted HERV-K113 element termed oriHERV-K113 using thin-section electron microscopy (EM) and could demonstrate that strong overexpression by substitution of the 5'LTR for a CMV promoter and partial codon optimization altered the virus assembly type and morphology. This included a conversion from the regular C-type to an A-type morphology with a mass of cytoplasmic immature cores tethered to the cell membrane and the membranes of vesicles. Overexpression permitted the release and maturation of virions but reduced the envelope content. A weaker boost of virus expression by Staufen-1 was not sufficient to induce these morphological alterations.
HERV-K(HML-2)家族包含人类基因组中最近整合且保存最完好的内源性前病毒序列。然而,所有已知元件都经历了突变或缺失,使得表达的颗粒无感染性。此外,这些插入后突变阻碍了对这个古老病毒家族一般生物学特性的分析。因此,共有序列以及具有回复插入后突变的元件序列的表达对于克服这一限制非常有帮助。我们使用超薄切片电子显微镜(EM)研究了一种最近重建的称为oriHERV-K113的HERV-K113元件的颗粒形态,结果表明,通过用巨细胞病毒(CMV)启动子替换5'长末端重复序列(LTR)以及部分密码子优化实现的强过表达改变了病毒组装类型和形态。这包括从规则的C型形态转变为A型形态,大量细胞质未成熟核心附着于细胞膜和囊泡膜上。过表达使病毒粒子得以释放和成熟,但降低了包膜含量。由Staufen-1介导的较弱的病毒表达增强不足以诱导这些形态改变。