Thymidine kinase (TK) activity in herpes simplex virus type 1 recombinants that carry insertions affecting regulation of the TK gene.
Thymidine kinase (TK) activity in herpes simplex virus type 1 recombinants that carry insertions affecting regulation of the TK gene.
复制标题
1 型单纯疱疹病毒重组体中的胸苷激酶 (TK) 活性，该重组体携带影响 TK 基因调节的插入。
DOI：
10.1016/0042-6822(86)90185-6
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发表时间：
1986
期刊：
影响因子：
3.7
通讯作者：
Edris,WA
中科院分区：
文献类型：
--
作者：
Tenser,RB;Edris,WA
确定单纯疱疹病毒1型（HSV）胸苷激酶（TK）表达在病毒潜伏期发病机制中的可能重要性部分取决于使用定义的突变体。在A. e.西尔斯，B。Meignier和B. Roizman（J. Virol.，55，410 - 416（1985）），其中他们利用了被认为是TK-的基因工程病毒重组体，据报道HSV TK表达在潜伏期中的作用是最小的。为了进一步研究这一结论，我们深入研究了其M316 - 2和M316 - 10 HSV-1突变体的TK表型。通过胸苷磷酸化、阿糖基胸腺嘧啶（ara-T）抑制和病毒空斑放射自显影研究TK活性。在5分钟试验中未检测到M316 - 2和M316 - 10 HSV突变体的TK活性（如Searset等人进行的），但在较长的测定中，明显具有实质性活性。相比之下，在对照TK −病毒的试验中，所有时间点的活性均极低或不存在。在ara-T抑制试验中，M316 - 2和M316 - 10病毒被抑制超过10倍，与中等TK活性的病毒一致。通过空斑放射自显影，这两种病毒都产生了含有大量胸苷的空斑。基于这些结果，我们得出结论，M316 - 2和M316 - 10病毒可能被认为表达中等水平的TK活性。使用这些突变体的HSV潜伏期结果可能需要考虑到这一点进行解释。
Determinations of the possible importance of herpes simplex virus type 1 (HSV) thymidine kinase (TK) expression in the pathogenesis of viral latency depend in part on the use of defined mutants. In a recent study by A. E. Sears, B. Meignier, and B. Roizman (J. Virol.,55, 410–416 (1985)), in which they utilized genetically engineered viral recombinants considered to be TK−, the role of HSV TK expression in latency was reported to be minimal. To further investigate this conclusion we intensively studied the TK phenotypes of their M316-2 and M316-10 HSV-1 mutants. TK activity was investigated by phosphorylation of thymidine, by arabinosylthymine (ara-T) inhibition and by virus plaque autoradiography. TK activity of the M316-2 and M316-10 HSV mutants was not detected in 5-min assays (as performed by Searset al.), but in longer assays substantial activity was apparent. In contrast, in assays of control TK−viruses, activity was minimal or absent at all time points. In ara-T inhibition assays the M316-2 and M316-10 viruses were inhibited more than 10-fold, consistent with viruses of intermediate TK activity. By plaque autoradiography both of these viruses produced plaques which incorporated significant amounts of thymidine. Based on these results we conclude that the M316-2 and M316-10 viruses should likely be considered to express intermediate levels of TK activity. HSV latency results using these mutants may need to be interpreted with this in mind.