Chou J, Roizman B
Marjorie B. Kovler Viral Oncology Laboratories, University of Chicago, Illinois 60637.
J Virol. 1990 Mar;64(3):1014-20. doi: 10.1128/JVI.64.3.1014-1020.1990.
In a previous study, it was reported that herpes simplex virus 1 (HSV-1) strain F contains a transcribed open reading frame situated in the inverted repeats of the L component between the terminal a sequence and the open reading frame that encodes the alpha 0 gene (J. Chou and B. Roizman, J. Virol. 57: 629-637, 1986). By means of an antibody to repeats of the trimer Ala-Thr-Pro predicted to be specified by the open reading frame, it was shown that the open reading frame specifies a protein (M. Ackermann, J. Chou, M. Sarmiento, R. A. Lerner, and B. Roizman, J. Virol. 58: 843-850, 1986). This open reading frame is absent from the reported sequence of HSV-1(17)syn+ (D. J. McGeoch, M. A. Dalrymple, A. J. Davison, A. Dolan, M. C. Frame, D. McNab, L. J. Perry, J. E. Scott, and P. Taylor, J. Gen. Virol. 69: 1531-1574, 1988; L. J. Perry and D. J. McGeoch, J. Gen. Virol. 69: 2831-2846, 1988). To define the extent of variability in this open reading frame, we compared the sequences of the ICP34.5-encoding open reading frames of the genomes of three strains characterized by limited passage in cell culture with that of the HSV-1(17)syn+ strain. Furthermore, to establish unambiguously that the antibody to the Ala-Thr-Pro repeats reacts with the product of this open reading frame, we inserted a short sequence that encodes a known epitope in frame at the 5' terminus of the coding domain. Our results indicate that with minor variations, the open reading frame is conserved in the three HSV-1 genomes analyzed but not in HSV-1(17)syn+. Thus, two strains contain an inserted amino acid and one strain, isolated from a case of human encephalitis, lacks a seven-amino-acid sequence. The recombinant virus carrying the foreign epitope expressed a slightly slower-migrating protein which reacted with both the rabbit polyclonal antibody to the Ala-Thr-Pro trimer repeats and the monoclonal antibody to the inserted epitope. The implications of the results are discussed.
在先前的一项研究中，据报道单纯疱疹病毒1型（HSV-1）F株含有一个转录的开放阅读框，该开放阅读框位于L组分的反向重复序列中，在末端a序列和编码α0基因的开放阅读框之间（J. 周和B. 罗兹曼，《病毒学杂志》57: 629 - 637, 1986年）。通过针对预计由该开放阅读框指定的三聚体Ala-Thr-Pro重复序列的抗体，表明该开放阅读框指定了一种蛋白质（M. 阿克曼、J. 周、M. 萨米恩托、R. A. 勒纳和B. 罗兹曼，《病毒学杂志》58: 843 - 850, 1986年）。HSV-1(17)syn+的报道序列中不存在这个开放阅读框（D. J. 麦吉奥克、M. A. 达尔林普尔、A. J. 戴维森、A. 多兰、M. C. 弗雷姆、D. 麦克纳布、L. J. 佩里、J. E. 斯科特和P. 泰勒，《普通病毒学杂志》69: 1531 - 1574, 1988年；L. J. 佩里和D. J. 麦吉奥克，《普通病毒学杂志》69: 2831 - 2846, 1988年）。为了确定这个开放阅读框的变异程度，我们将三种在细胞培养中传代有限的毒株基因组中编码ICP34.5的开放阅读框序列与HSV-1(17)syn+毒株的序列进行了比较。此外，为了明确证明针对Ala-Thr-Pro重复序列的抗体与这个开放阅读框的产物发生反应，我们在编码结构域的5'末端框内插入了一个编码已知表位的短序列。我们的结果表明，虽有微小差异，但在所分析的三个HSV-1基因组中该开放阅读框是保守的，而在HSV-1(17)syn+中则不然。因此，两个毒株含有一个插入的氨基酸，而从一例人类脑炎病例中分离出的一个毒株缺少一个七氨基酸序列。携带外源表位的重组病毒表达了一种迁移速度稍慢的蛋白质，它与针对Ala-Thr-Pro三聚体重复序列的兔多克隆抗体以及针对插入表位的单克隆抗体都发生反应。文中讨论了这些结果的意义。
