Marchetti M E, Smith C A, Schaffer P A
Laboratory of Tumor Virus Genetics, Dana-Farber Cancer Institute, Boston, Massachusetts.
J Virol. 1988 Mar;62(3):715-21. doi: 10.1128/JVI.62.3.715-721.1988.
ts701 is a temperature-sensitive mutant of herpes simplex virus type 1 strain KOS induced by hydroxylamine mutagenesis (C.T. Chu, D. S. Parris, R. A. F. Dixon, F. E. Farber, and P. A. Schaffer, Virology 98:168-181, 1979). In the present study, the mutation rendering ts701 temperature sensitive was mapped to coordinates 0.609 through 0.614 in the UL region of the genome. At the nonpermissive temperature, ts701 (i) failed to induce the synthesis of viral DNA, (ii) exhibited a dramatically reduced ability to drive replication of a plasmid containing the herpes simplex virus origin of viral DNA synthesis, oriS, (iii) generated no viral polypeptides of the late (gamma 2) kinetic class, and (iv) produced virions with electron-translucent cores. Northern (RNA) blot hybridization demonstrated that two mRNAs--one of the beta kinetic class and one of the gamma kinetic class--hybridized to a 1.3-kilobase viral DNA fragment that rescued the mutation in ts701. Based on the phenotype and mapping of ts701, it is likely that its mutation lies in the gene specifying the 65,000-Mr DNA-binding protein (65KDBP) recently described by Marsden et al. (H.S. Marsden, M.E.M. Campbell, L. Haarr, M. C. Frame, D. S. Parris, M. Murphy, R. G. Hope, M. T. Muller, and C. M. Preston, J. Virol. 61:2428-2437, 1987).
ts701是通过羟胺诱变产生的单纯疱疹病毒1型KOS株的温度敏感突变体（C.T.朱、D.S.帕里斯、R.A.F.迪克森、F.E.法伯和P.A.沙弗，《病毒学》98:168 - 181，1979）。在本研究中，使ts701具有温度敏感性的突变被定位到基因组UL区域中坐标为0.609至0.614的位置。在非允许温度下，ts701：（i）未能诱导病毒DNA的合成；（ii）驱动含有单纯疱疹病毒病毒DNA合成起始点oriS的质粒复制的能力显著降低；（iii）未产生晚期（γ2）动力学类别的病毒多肽；（iv）产生具有电子透明核心的病毒粒子。Northern（RNA）印迹杂交表明，两种mRNA（一种是β动力学类别，一种是γ动力学类别）与一个1.3千碱基的病毒DNA片段杂交，该片段挽救了ts701中的突变。基于ts701的表型和定位，其突变可能位于最近由马斯登等人描述的指定65,000道尔顿DNA结合蛋白（65KDBP）的基因中（H.S.马斯登、M.E.M.坎贝尔、L.哈阿尔、M.C.弗雷姆、D.S.帕里斯、M.墨菲、R.G.霍普、M.T.穆勒和C.M.普雷斯顿，《病毒学杂志》61:2428 - 2437，1987）。
