Bloss T A, Sugden B
McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
J Virol. 1994 Dec;68(12):8217-22. doi: 10.1128/JVI.68.12.8217-8222.1994.
We measured the efficiency of DNA packaging by Epstein-Barr virus (EBV) as a function of the length of the DNA being packaged. Plasmids that contain oriP (the origin of latent EBV DNA replication), oriLyt (the origin of lytic EBV DNA replication), the viral terminal repeats (necessary for cleavage and packaging by EBV), and various lengths of bacteriophage lambda DNA were introduced into EBV-positive cells. Upon induction of the resident EBV's lytic phase, introduced plasmids replicated as concatemers and were packaged. Plasmid-derived concatemers of DNA with certain lengths were found to predominate in isolated virion particles. We measured the distribution of lengths of plasmid concatemers found within cells supporting the lytic phase of the viral life cycle and found that this distribution differed from the distribution of lengths of concatemers found in mature virion particles. This finding indicates that the DNA packaged into mature virions represents a selected subset of those present in the cell during packaging. These observations together indicate that the length of DNA affects the efficiency with which that DNA is packaged by EBV. Finally, we measured the length of the packaged B95-8 viral DNA and found it to be approximately 165 kbp, or 10 kbp shorter than the originally predicted size for B95-8 based on its sequence. Together with the results of other studies, these findings indicate that the packaging of DNAs by EBV is dependent on two imprecisely recognized elements: the viral terminal repeats and the length of the DNA being packaged by the virus.
我们测量了爱泼斯坦-巴尔病毒(EBV)包装DNA的效率与被包装DNA长度之间的函数关系。将含有oriP(EBV潜伏性DNA复制起点)、oriLyt(EBV裂解性DNA复制起点)、病毒末端重复序列(EBV进行切割和包装所必需)以及不同长度噬菌体λDNA的质粒导入EBV阳性细胞。在诱导宿主EBV进入裂解期后,导入的质粒以多联体形式复制并被包装。发现具有特定长度的质粒衍生DNA多联体在分离出的病毒粒子中占主导地位。我们测量了在支持病毒生命周期裂解期的细胞内发现的质粒多联体的长度分布,发现该分布与在成熟病毒粒子中发现的多联体长度分布不同。这一发现表明,包装到成熟病毒粒子中的DNA代表了包装过程中细胞内存在的DNA的一个选定子集。这些观察结果共同表明,DNA的长度会影响EBV对其包装的效率。最后,我们测量了被包装的B95-8病毒DNA的长度,发现其约为165 kbp,比根据其序列最初预测的B95-8大小短10 kbp。与其他研究结果一起,这些发现表明EBV对DNA的包装依赖于两个识别不精确的元件:病毒末端重复序列和病毒所包装DNA的长度。